RESUMEN

OBJECTIVE: This study aimed to evaluate neuronal markers in stromal cells from human exfoliated deciduous teeth (SHED) and standardize the isolation and characterization of those cells. METHODOLOGY: Healthy primary teeth were collected from children. The cells were isolated by enzymatic digestion with collagenase. By following the International Society for Cell and Gene Therapy (ISCT) guidelines, SHED were characterized by flow cytometry and differentiated into osteogenic, adipogenic, and chondrogenic lineages. Colony-forming unit-fibroblasts (CFU-F) were performed to assess these cells' potential and efficiency. To clarify the neuronal potential of SHED, the expression of nestin and ßIII-tubulin were examined by immunofluorescence and SOX1, SOX2, GFAP, and doublecortin (DCX), nestin, CD56, and CD146 by flow cytometry. RESULTS: SHED showed mesenchymal stromal cells characteristics, such as adhesion to plastic, positive immunophenotypic profile for CD29, CD44, CD73, CD90, CD105, and CD166 markers, reduced expression for CD14, CD19, CD34, CD45, HLA-DR, and differentiation in three lineages confirmed by staining and gene expression for adipogenic differentiation. The average efficiency of colony formation was 16.69%. SHED expressed the neuronal markers nestin and ßIII-tubulin; the fluorescent signal intensity was significantly higher in ßIII-tubulin (p<0.0001) compared to nestin. Moreover, SHED expressed DCX, GFAP, nestin, SOX1, SOX2, CD56, CD146, and CD271. Therefore, SHED had a potential for neuronal lineage even without induction with culture medium and specific factors. CONCLUSION: SHEDs may be a new therapeutic strategy for regenerating and repairing neuronal cells and tissues.

Asunto(s)

Células Madre Mesenquimatosas , Tubulina (Proteína) , Niño , Humanos , Nestina/metabolismo , Tubulina (Proteína)/metabolismo , Antígeno CD146/metabolismo , Diferenciación Celular/fisiología , Células Madre Mesenquimatosas/metabolismo , Diente Primario , Células Cultivadas , Células del Estroma

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INTRODUCTION: Endothelial progenitor cells (EPCs) and nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme activity may affect the vessel wall and have a role in development of aortic aneurysms. EPCs originate from hematopoietic stem cells and can be enumerated from peripheral blood samples by flow cytometry. In this study, we aimed to evaluate the relation of EPC number and NADPH oxidase enzyme activity in the development of thoracic aortic aneurysm (TAA). METHODS: Patients with TAA (n=30) and healthy individuals without TAA (control, n=10) were included in our study. Characterization and enumeration of EPC from peripheral blood samples were performed by flow cytometry with panels including markers of EPCs (CD34/CD133/CD309/CD146/CD144). Additionally, NADPH oxidase enzyme activity (capacity) was also measured by the dihydrorhodamine 123 (DHR 123) test. RESULTS: The enumeration of EPC with CD34+/CD146+ marker showed that the number of mean EPC/106 cells was increased in the patient group (41.5/106 cells), but not in the control group (20.50/105 cells) (P<0.01). Additionally, patients with TAA presented significantly lower NADPH oxidase activity by DHR assay than healthy controls (mean stimulation index: 60.40± 7.86 and 75.10±5.21, respectively) (P<0.01). CONCLUSION: Our results showed that the number of EPCs is significantly higher in aortic aneurysm patients and may have a role in disease progression. The crosstalk between NADPH oxidase enzyme capacity and EPC number may be useful as a parameter to explain the clinical progression of TAA.

Asunto(s)

Aneurisma de la Aorta , Células Progenitoras Endoteliales , Antígenos CD34 , Biomarcadores , Antígeno CD146 , Humanos , NADPH Oxidasas , Células Madre

RESUMEN

Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. OBJECTIVE: Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. METHODOLOGY: The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. RESULTS: Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. CONCLUSIONS: These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Asunto(s)

Células Madre Mesenquimatosas , Odontogénesis , Animales , Antígeno CD146 , Diferenciación Celular , Ratones , Ratones Endogámicos BALB C , Receptores de Factor de Crecimiento Nervioso , Células Madre

RESUMEN

Abstract Mesenchymal and epithelial stem cells were identified in dental tissues; however, knowledge about the odontogenic stem cells is limited, and there are some questions regarding their temporo-spatial dynamics in tooth development. Objective Our study aimed to analyze the expression of the stem cell markers CD146 and p75NTR during the different stages of odontogenesis. Methodology The groups consisted of 13.5, 15.5, 17.5 days old embryos, and 14 days postnatal BALB/c mice. The expression of CD146 and p75NTR was evaluated by immunohistochemistry. Results Our results showed that positive cells for both markers were present in all stages of tooth development, and the number of positive cells increased with the progression of this process. Cells of epithelial and ectomesenchymal origin were positive for CD146, and the expression of p75NTR was mainly detected in the dental papilla and dental follicle. In the postnatal group, dental pulp cells were positive for CD146, and the reduced enamel epithelium and the oral mucosa epithelium showed immunostaining for p75NTR. Conclusions These results suggest that the staining pattern of CD146 and p75NTR underwent temporal and spatial changes during odontogenesis and both markers were expressed by epithelial and mesenchymal cell types, which is relevant due to the significance of the epithelial-ectomesenchymal interactions in tooth development.

Asunto(s)

Animales , Ratones , Células Madre Mesenquimatosas , Odontogénesis , Células Madre , Diferenciación Celular , Receptores de Factor de Crecimiento Nervioso , Antígeno CD146 , Ratones Endogámicos BALB C

RESUMEN

Dental stem cells have many applications in medicine, dentistry and stem cell biology in general due to their easy accessibility and low morbidity. A common surgical manoeuvre after a tooth extraction is the dental socket curettage which is necessary to clean the alveolus and favour alveolar bone healing. This procedure can cause very low morbidity compared to bone marrow collection procedures and the collected material is normally discarded. In order to investigate if the tissue obtained by dental socket curettage after a tooth extraction was a feasible alternative source to isolate human stem cells, we isolated and characterized two different stem cell populations based on STRO-1 and CD146 expression. We were able to collect and grow cells from dental socket of vital and non-vital teeth. Both populations were proliferative, clonogenic and expressed STRO-1, CD146, CD90, NG2, PDGFR-ß, which are markers found in stem cells, presented in vitro multiline-differentiation into osteogenic, chondrogenic, and adipogenic tissue, and in vivo transplanted cells formed mineralized tissue. Interestingly, STRO-1+ clonogenic cells presented better multidifferentiation than CD146+ cells. Our results showed that mesenchymal stem cells can be isolated from the tiny tissue collected by dental socket curettage after vital and non-vital tooth extraction and suggest that STRO-1 is an important marker to be used to sort cells with multidifferentiation capacity.

Asunto(s)

Diferenciación Celular , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Alveolo Dental/citología , Animales , Antígenos de Superficie/análisis , Antígeno CD146/análisis , Proliferación Celular , Células Cultivadas , Humanos , Separación Inmunomagnética , Masculino , Ratones Endogámicos BALB C , Ratones Desnudos

RESUMEN

Duchenne muscular dystrophy is the most common and severe form of progressive muscular dystrophy. Previous results showed an increased survival in double knockout mice (dko) when treated with adipose-derived CD146+ cells. In this study, we analyzed the effect of CD146+ cells compared to mesenchymal stem/stromal cells (MSCs) derived from the same human adipose sample when injected in the dko mouse model without immunosuppression. Both CD146+ cells and MSCs increased the survival of treated mice when compared to vehicle-injected mice, with a more prominent effect of CD146+ cells than MSCs. Both CD146+ cells and MSCs suppressed peripheral blood mononuclear cell proliferation, indicating immunomodulatory properties. Co-culture experiments showed that MSCs have a more inflammatory profile expression, and angiogenesis assay showed that CD146+ cells can improve blood vessel formation. CD146+ cells can extend survival of muscular dystrophy mice more efficiently than MSCs, possibly due to immunomodulatory and angiogenic properties. Further investigations focusing on exogenous CD146+ cell role in vivo will improve cell therapy understanding and effectiveness.

Asunto(s)

Adipocitos/citología , Antígeno CD146/metabolismo , Modelos Animales de Enfermedad , Células Madre Mesenquimatosas/citología , Distrofia Muscular Animal/terapia , Neovascularización Fisiológica , Adipocitos/metabolismo , Animales , Diferenciación Celular , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Células Endoteliales de la Vena Umbilical Humana/citología , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/metabolismo , Células Madre Mesenquimatosas/metabolismo , Ratones , Ratones SCID , Distrofia Muscular Animal/metabolismo , Distrofia Muscular Animal/patología

RESUMEN

Endometriosis is a chronic gynecological disorder defined as the presence of endometrial tissue within extra-uterine sites. The primary symptoms are infertility and chronic pain. The inflammatory environment and aberrant immune responses in women with endometriosis may be directly associated with the initiation and progression of endometriotic lesions. In the present study, the secretion of inflammatory cytokines was evaluated in cultures of primary endometrial cells (ECs) isolated from the endometrium of women with and without endometriosis. The presence of endometriotic cells leads to alterations in the secretory profile of healthy ECs. The expression of the inflammatory cytokines interleukin (IL)­6 and IL­8 was significantly increased in endometriotic and co­cultured cells compared with healthy ECs. IL­6 expression was strongly correlated with IL­8 expression in endometriotic cells. IL­1ß expression was increased on day 10 of co­culture to 48.30 pg/ml and may be associated with the long­term co­culture, rather than IL­6 and IL­8 expression. IL­6 expression was strongly correlated with cell number, whereas IL­8 expression was moderately correlated with cell number. Additionally, it was observed that co­cultured cells exhibited a different population of cells, with expression of the mesenchymal stem cell marker cell surface glycoprotein MUC18, indicating a putative role of endometrial mesenchymal stem cells in the secretion of cytokines and disease development. These results indicate a predominant role of primary endometriotic cells in the secretion of cytokines, which contributes to the disrupted peritoneal and endometrial environment observed in the women with endometriosis.

Asunto(s)

Endometriosis/metabolismo , Regulación de la Expresión Génica , Interleucina-6/biosíntesis , Interleucina-8/biosíntesis , Adolescente , Adulto , Antígeno CD146 , Técnicas de Cocultivo , Endometriosis/patología , Femenino , Perfilación de la Expresión Génica , Humanos , Inflamación/metabolismo , Inflamación/patología , Persona de Mediana Edad

RESUMEN

We investigated for the first time the expression of melanoma cell adhesion molecule (MCAM) and its involvement in the differentiation of 3T3-L1 fibroblasts to adipocytes. We found that MCAM mRNA increased subsequent to the activation of the master regulator of adipogenesis, PPARγ, and this increase was maintained in the mature adipocytes. On the other hand, MCAM knockdown impaired differentiation and induction of PPARγ as well as expression of genes activated by PPARγ. However, events that precede and are necessary for early PPARγ activation, such as C/EBPß induction, ß-catenin downregulation, and ERK activation, were not affected in the MCAM knockdown cells. In keeping with this, the increase in PPARγ mRNA that precedes MCAM induction was not altered in the knockdown cells. In conclusion, our findings suggest that MCAM is a gene upregulated and involved in maintaining PPARγ induction in the late but not in the early stages of 3T3-L1 fibroblasts adipogenesis.

Asunto(s)

Adipocitos/metabolismo , Adipogénesis , Diferenciación Celular , Fibroblastos/metabolismo , Regulación de la Expresión Génica , PPAR gamma/biosíntesis , Células 3T3-L1 , Adipocitos/citología , Animales , Antígeno CD146/genética , Antígeno CD146/metabolismo , Fibroblastos/citología , Técnicas de Silenciamiento del Gen , Ratones , PPAR gamma/genética

RESUMEN

BACKGROUND: Patients with breast cancer-the deadliest cancer among women-are at constant risk of developing metastasis. Oxidative stress and hypoxia are common feature of tumor cells that can proliferate even in a resultant metabolic acidosis. Despite the low extracellular pH, intracellular pH of tumor cells remains relatively normal, or even more alkaline due to the action of a membrane protein family known as monocarboxylate transporters (MCTs). The objective of this study was to verify the diagnostic and prognostic value of MCT1, MCT4 and CD147 in tumor and peripheral blood samples of patients with breast cancer undergoing chemotherapic treatment. METHODS: Differential expression of MCT1, MCT4 and CD147 obtained by qPCR was determined by 2-ΔΔCq method between biological samples (tumor and serial samples of peripheral) of patients (n = 125) and healthy women (n = 25). RESULTS: tumor samples with higher histological grades have shown higher expression of these markers; this higher expression was also observed in blood samples obtained at diagnosis of patients when compared to healthy women and in patients with positive progression of the disease (metastasis development). CONCLUSION: markers studied here could be a promising strategy in routine laboratory evaluations as breast cancer diagnosis and prognosis.

Asunto(s)

Biomarcadores de Tumor/sangre , Neoplasias de la Mama/sangre , Antígeno CD146/sangre , Transportadores de Ácidos Monocarboxílicos/sangre , Proteínas Musculares/sangre , Simportadores/sangre , Adulto , Anciano , Estudios de Casos y Controles , Femenino , Humanos , Persona de Mediana Edad

RESUMEN

UNLABELLED: Angiogenesis is a key process for metastatic progression. While it has been established that the evaluation of breast tumoral microvessel density by CD105 marker is a potential prognostic parameter, its evaluation by CD146 marker has been poorly studied. AIM: The purpose of this study was to compare the prognostic value of intra-tumoral microvessel density assayed by CD105 and CD146 in early breast cancer patients. METHODS: 42 women with breast infiltrative ductal carcinoma (I and II-stages) were retrospectively reviewed. Intra-tumoral microvessel density was immunohistochemically examined using antibodies anti-CD105 and CD146 in paraffin-embedded tissues, and their association with classical prognostic-markers, metastatic recurrence, metastasis-free survival and overall survival was analyzed. RESULTS: High microvessel density assessed by CD146 was significantly associated with a higher risk of developing metastasis (p=0.0310) and a shorter metastasis-free survival (p=0.0197). In contrast, when we used the CD105-antibody, we did not find any significant association. Finally, CD146 showed to be an independent predictive indicator for metastasis-free survival (p=0.0055). CONCLUSION: Our data suggest that the intra-tumoral microvessel density evaluated by CD146 may be a more suitable predictor of metastatic development than that evaluated by CD105 in early breast cancer.

Asunto(s)

Biomarcadores de Tumor/análisis , Neoplasias de la Mama/irrigación sanguínea , Carcinoma Ductal de Mama/irrigación sanguínea , Endoglina/análisis , Adulto , Neoplasias de la Mama/mortalidad , Neoplasias de la Mama/patología , Antígeno CD146/análisis , Carcinoma Ductal de Mama/mortalidad , Carcinoma Ductal de Mama/patología , Femenino , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Persona de Mediana Edad , Neovascularización Patológica/patología , Proyectos Piloto , Pronóstico , Modelos de Riesgos Proporcionales , Estudios Retrospectivos

RESUMEN

OBJECTIVE: Previous work demonstrated the effectiveness of autologous adipose-derived stem cells (ASCs) as endothelial cell (EC) substitutes in vascular tissue engineering. We further this work toward clinical translation by evaluating ASC function after (1) replacement of fetal bovine serum (FBS) with autologous human plasma (HP) in culture and (2) cryopreservation. METHODS: Human ASCs and plasma, isolated from periumbilical fat and peripheral blood, respectively, were collected from the same donors. ASCs were differentiated in endothelial growth medium supplemented with FBS (2%) vs HP (2%). Proliferation was measured by growth curves and MTT assay. Endothelial differentiation was measured by quantitative polymerase chain reaction, assessment of acetylated low-density lipoprotein uptake, and cord formation after plating on Matrigel (BD Biosciences, San Jose, Calif). Similar studies were conducted before and after cryopreservation of ASCs and included assessment of cell retention on the luminal surface of a vascular graft. RESULTS: ASCs expanded in HP-supplemented medium showed (1) similar proliferation to FBS-cultured ASCs, (2) consistent differentiation toward an EC lineage (increases in CD31, von Willebrand factor, and CD144 message; acetylated low-density lipoprotein uptake; and cord formation on Matrigel), and (3) retention on the luminal surface after seeding and subsequent flow conditioning. Cryopreservation did not significantly alter ASC viability, proliferation, acquisition of endothelial characteristics, or retention after seeding onto a vascular graft. CONCLUSIONS: This study suggests that (1) replacement of FBS with autologous HP--a step necessary for the translation of this technology into human use--does not significantly impair proliferation or endothelial differentiation of ASCs used as EC substitutes and (2) ASCs are tolerant to cryopreservation in terms of maintaining EC characteristics and retention on a vascular graft.

Asunto(s)

Tejido Adiposo/citología , Bioprótesis , Prótesis Vascular , Técnicas de Cultivo de Célula , Criopreservación , Células Progenitoras Endoteliales/metabolismo , Plasma/metabolismo , Ingeniería de Tejidos/métodos , Biomarcadores/metabolismo , Antígeno CD146/genética , Antígeno CD146/metabolismo , Diferenciación Celular , Linaje de la Célula , Proliferación Celular , Supervivencia Celular , Células Cultivadas , Humanos , Lipoproteínas LDL/metabolismo , Neovascularización Fisiológica , Fenotipo , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/genética , Molécula-1 de Adhesión Celular Endotelial de Plaqueta/metabolismo , Factores de Tiempo , Factor de von Willebrand/genética , Factor de von Willebrand/metabolismo

RESUMEN

Several studies have confirmed that the breast tumor microenvironment drives cancer progression and metastatic development. The aim of our research was to investigate the prognostic significance of the breast tumor microenvironment in untreated early breast cancer patients. Therefore, we analyzed the association of the expression of α-SMA, FSP, CD105 and CD146 in CD34-negative spindle-shaped stromal cells, not associated with the vasculature, in primary breast tumors with classical prognostic marker levels, metastatic recurrence, local relapse, disease-free survival, metastasis-free survival and the overall survival of patients. In the same way, we evaluated the association of the amount of intra-tumor stroma, fibroblasts, collagen deposition, lymphocytic infiltration and myxoid changes in these samples with the clinical-pathological data previously described. This study is the first to demonstrate the high CD105 expression in this stromal cell type as a possible independent marker of unfavorable prognosis in early breast cancer patients. Our study suggests that this new finding can be useful prognostic marker in the clinical-pathological routine.

Asunto(s)

Antígenos CD34/metabolismo , Antígenos CD/metabolismo , Biomarcadores de Tumor/metabolismo , Neoplasias de la Mama/metabolismo , Neoplasias de la Mama/patología , Regulación Neoplásica de la Expresión Génica , Receptores de Superficie Celular/metabolismo , Actinas/metabolismo , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias de la Mama/diagnóstico , Antígeno CD146/metabolismo , Supervivencia sin Enfermedad , Endoglina , Femenino , Humanos , Persona de Mediana Edad , Metástasis de la Neoplasia , Recurrencia , Estudios Retrospectivos , Células del Estroma/metabolismo , Células del Estroma/patología , Microambiente Tumoral

RESUMEN

Changes in oxygen concentration may influence various innate characteristics of stem cells. The effects of varying oxygen concentration on human periodontal ligament stem cells (HPDLSCs) has not been explored, particularly under hypoxia-related conditions. First, HPDLSCs were cultured from the periodontium of human teeth using the outgrowth method. STRO-1 and CD146 expression of HPDLSCs was investigated by flow cytometry. To detect the multilineage differentiation capacities of HPDLSCs, osteogenic-like and adipogenic-like states were induced in cells. Next, HPDLSCs (passage 3) were exposed to normal oxygen (21% O2) or hypoxia (2% O2) conditions for 7 days and cell proliferation was evaluated. After culture in osteogenic medium for 7 days, osteoblastic differentiation was evaluated by semi-quantitative reverse transcription-polymerase chain reaction analysis to detect 3 osteoblastic markers: core-binding factor a 1/runt-related transcription factor 2, osteocalcin, and osteopontin. In addition, each cell group was incubated with a hydroxyapatite/tricalcium phosphate carrier and transplanted subcutaneously into the back of immunocompromised mice to investigate transplantation differences in vivo. HPDLSCs were isolated, cultured, and successfully identified. After exposure of HPDLSCs to hypoxia for 7 days, the proliferation rate was increased and showed higher osteogenic differentiation potential compared to control cells. After 12 weeks of transplantation, hypoxia-treated HPDLSCs differentiated into osteoblast-like cells that formed bone-like structures. These results suggest that oxygen concentrations affect various aspects of HPDLSC physiology and that hypoxia enhances osteogenic differentiation both in vivo and in vitro. Oxygen concentration may be a critical parameter for HPDLSCs during expansion and differentiation.

Asunto(s)

Técnicas de Cultivo de Célula/métodos , Osteogénesis , Ligamento Periodontal/citología , Ligamento Periodontal/metabolismo , Células Madre/citología , Adolescente , Animales , Antígenos de Superficie/metabolismo , Biomarcadores , Antígeno CD146/metabolismo , Diferenciación Celular , Hipoxia de la Célula , Proliferación Celular , Células Cultivadas , Medios de Cultivo/química , Humanos , Ratones , Trasplante de Células Madre , Células Madre/metabolismo , Adulto Joven

RESUMEN

INTRODUCTION: Previous studies describe contrasting molecular profiles of active and inactive periapical granulomas characterized by distinct expression of cytokines, osteoclastogenic factors, and wound healing markers. Although the molecular mechanisms underlying such a dichotomy remain unknown, in this study we investigated the potential involvement of mesenchymal stem cells (MSCs) in determining human and murine periapical lesion activity and outcomes. METHODS: Periapical granulomas (n = 83) and control samples (n = 24) were comparatively assessed for the expression levels of 11 mesenchymal stem cell (MSC) markers using real-time polymerase chain reaction. Experimental periapical lesions induced in mice were evaluated for MSC marker expression and the effects of AMD3100 treatment on lesion outcomes. RESULTS: MCS marker expression was prevalent in periapical granulomas compared with that in controls, whereas CD29, CD73, CD90, CD146, CD166, NANOG, Stro-1, and CXCR4 expressions were higher in inactive than in active lesions. Experimental periapical lesion inactivity was also associated with an increased expression of MSC markers. The inhibition of MSC mobilization to the periapex by AMD3100 resulted in increased lesion sizes; decreased expression of MSCs and wound healing markers; and increased expression of interleukin 1 beta (IL-17ß), interleukin 17 (IL-17), tumor necrosis factor alpha (TNF-α), interferon gamma (IFN-γ), and nuclear factor kappa-B ligand (RANKL). CONCLUSIONS: Our results show that MSC markers are overexpressed in inactive human and experimental periapical lesions and that MSC mobilization results in the attenuation of experimental lesion progression associated with immunosuppressive and prohealing mechanisms.

Asunto(s)

Inmunosupresores/farmacología , Células Madre Mesenquimatosas/fisiología , Granuloma Periapical/patología , 5'-Nucleotidasa/análisis , Molécula de Adhesión Celular del Leucocito Activado/análisis , Adulto , Animales , Antígenos de Superficie/análisis , Bencilaminas , Biomarcadores/análisis , Antígeno CD146/análisis , Ciclamas , Modelos Animales de Enfermedad , Compuestos Heterocíclicos/uso terapéutico , Proteínas de Homeodominio/análisis , Humanos , Integrina beta1/análisis , Interferón gamma/análisis , Interleucina-17/análisis , Interleucina-1beta/análisis , Células Madre Mesenquimatosas/efectos de los fármacos , Ratones , Persona de Mediana Edad , Granuloma Periapical/tratamiento farmacológico , Granuloma Periapical/fisiopatología , Tejido Periapical/citología , Tejido Periapical/efectos de los fármacos , Tejido Periapical/fisiología , Ligando RANK/análisis , Receptores CXCR4/análisis , Receptores CXCR4/antagonistas & inhibidores , Antígenos Thy-1/análisis , Factor de Necrosis Tumoral alfa/análisis , Cicatrización de Heridas/fisiología

RESUMEN

BACKGROUND: MCAM has been recently identified as a biomarker for epithelial-mesenchymal transition (EMT) and is potentially involved in metastasis of cancer. The current study aimed at investigating the expression of MCAM in non-small-cell lung cancer (NSCLC) and its clinico-pathological significance. METHODS: A follow-up analysis was performed on 118 patients with NSCLC resected by lobectomy or pneumectomy with systematic lymph node dissection. All patients were followed for 6-60 months. Immunostaining of tissue sections from primary tumors and their lymph node metastasis was performed and evaluated using monoclonal antibody against MCAM, E-cadherin, and vimentin. Correlations were investigated between MCAM immunostaining in primary tumors and E-cadherin, vimentin immunostaining, lymph node metastasis, and survival. RESULTS: MCAM protein expression was found in 46.61 % of squamous cell carcinomas and 37.47 % of adenocarcinomas; MCAM expression positively correlated with vimentin, but inversely with E-cadherin (both P values <0.05). There were significant correlations between the MCAM immunostaining score in primary tumors and in their lymph node metastasis (P = 0.03). According to the Kaplan-Meier survival estimate, the level of MCAM expression in primary tumors was a statistically significant prognostic factor (P < 0.05). CONCLUSIONS: MCAM expression in surgically treated NSCLC is clearly associated with lymph node metastasis and poor prognosis.

Asunto(s)

Carcinoma de Pulmón de Células no Pequeñas/diagnóstico , Carcinoma de Pulmón de Células no Pequeñas/mortalidad , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/mortalidad , Adulto , Anciano , Biomarcadores de Tumor/metabolismo , Antígeno CD146/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/metabolismo , Carcinoma de Pulmón de Células no Pequeñas/patología , Femenino , Humanos , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patología , Metástasis Linfática , Masculino , Persona de Mediana Edad , Pronóstico , Estudios Retrospectivos , Análisis de Supervivencia

RESUMEN

Bone marrow stromal cells (BMSCs) are a valuable resource for skeletal regenerative medicine because of their osteogenic potential. In spite of the very general term "stem cell," this population of cells is far from homogeneous, and different BMSCs clones have greatly different phenotypic properties and, therefore, potentially different therapeutic potential. Adherence to a culture flask surface is a primary defining characteristic of BMSCs. We hypothesized that based on the adherence time we could obtain an enriched population of cells with a greater therapeutic potential. We characterized two populations of bone marrow-derived cells, those that adhered by three days (R-cells) and those that did not adhere by three days but did by six days (L-cells). Clones derived from L-cells could be induced into adipogenic, chondrogenic, and osteogenic differentiation in vitro. L-cells appeared to have greater proliferative capacity, as manifested by larger colony diameter and clones with higher CD146 expression. Only clones from L-cells developed bone marrow stroma in vivo. We conclude that the use of late adherence of BMSCs is one parameter that can be used to enrich for cells that will constitute a superior final product for cell therapy in orthopedics.

Asunto(s)

Adhesión Celular/genética , Diferenciación Celular , Células Madre Mesenquimatosas/citología , Osteogénesis , Nicho de Células Madre , Adulto , Antígeno CD146/biosíntesis , Linaje de la Célula/genética , Células Cultivadas , Femenino , Fibroblastos/citología , Regulación de la Expresión Génica , Humanos , Masculino , Persona de Mediana Edad , Medicina Regenerativa

RESUMEN

OBJECTIVE: The relationship of multipotent mesenchymal stromal cells (MSC) with pericytes and fibroblasts has not been established thus far, although they share many markers of primitive marrow stromal cells and the osteogenic, adipogenic, and chondrogenic differentiation potentials. MATERIALS AND METHODS: We compared MSCs from adult or fetal tissues, MSC differentiated in vitro, fibroblasts and cultures of retinal pericytes obtained either by separation with anti-CD146 or adhesion. The characterizations included morphological, immunophenotypic, gene-expression profile, and differentiation potential. RESULTS: Osteogenic, adipocytic, and chondrocytic differentiation was demonstrated for MSC, retinal perivascular cells, and fibroblasts. Cell morphology and the phenotypes defined by 22 markers were very similar. Analysis of the global gene expression obtained by serial analysis of gene expression for 17 libraries and by reverse transcription polymerase chain reaction of 39 selected genes from 31 different cell cultures, revealed similarities among MSC, retinal perivascular cells, and hepatic stellate cells. Despite this overall similarity, there was a heterogeneous expression of genes related to angiogenesis, in MSC derived from veins, artery, perivascular cells, and fibroblasts. Evaluation of typical pericyte and MSC transcripts, such as NG2, CD146, CD271, and CD140B on CD146 selected perivascular cells and MSC by real-time polymerase chain reaction confirm the relationship between these two cell types. Furthermore, the inverse correlation between fibroblast-specific protein-1 and CD146 transcripts observed on pericytes, MSC, and fibroblasts highlight their potential use as markers of this differentiation pathway. CONCLUSION: Our results indicate that human MSC and pericytes are similar cells located in the wall of the vasculature, where they function as cell sources for repair and tissue maintenance, whereas fibroblasts are more differentiated cells with more restricted differentiation potential.

Asunto(s)

Antígeno CD146/genética , Fibroblastos/citología , Perfilación de la Expresión Génica , Células Madre Mesenquimatosas/citología , Pericitos/citología , Cordón Umbilical/citología , Antígeno CD146/fisiología , Diferenciación Celular/fisiología , Separación Celular/métodos , Células Cultivadas , Análisis por Conglomerados , Fibroblastos/fisiología , Citometría de Flujo , Humanos , Inmunofenotipificación , Células Madre Mesenquimatosas/fisiología , Pericitos/fisiología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética/genética , Cordón Umbilical/fisiología

RESUMEN

The association between breast cancer initiation and prolonged exposure to estrogen suggests that this hormone may also have an etiologic role in such a process. On the other hand, many studies have found an association between human cancer and exposure to agricultural pesticides such as parathion, an organophosphorous pesticide used in agriculture to control mosquito plagues. However, the key factors behind the initiation of breast cancer remain to be elucidated. The aim of this study was to determine the effect of 17beta estradiol (estrogen) and parathion on protein expression in cell transformation of human breast epithelial cells in vitro. Estrogen and parathion alone and in combination induced malignant transformation of an immortalized human breast epithelial cell line, MCF-I0F, as indicated by anchorage independency and invasive capabilities. The results indicate that a combination of estrogen and parathion increased the expression of related cell adhesion proteins such as Dvl, Notch, CD146 and beta catenin. In conclusion, it can be suggested that pesticides affect human breast cell adhesion changes indicative of transformation.

Asunto(s)

Mama/efectos de los fármacos , Moléculas de Adhesión Celular/efectos de los fármacos , Células Epiteliales/efectos de los fármacos , Estradiol/toxicidad , Insecticidas/toxicidad , Paratión/toxicidad , Proteínas Adaptadoras Transductoras de Señales/efectos de los fármacos , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Mama/metabolismo , Antígeno CD146/efectos de los fármacos , Antígeno CD146/metabolismo , Línea Celular Tumoral , Transformación Celular Neoplásica/efectos de los fármacos , Proteínas Dishevelled , Femenino , Humanos , Inmunohistoquímica , Fosfoproteínas/efectos de los fármacos , Fosfoproteínas/metabolismo , Receptores Notch/efectos de los fármacos , Receptores Notch/metabolismo , beta Catenina/efectos de los fármacos , beta Catenina/metabolismo

RESUMEN

Mel-CAM (CD146) is a cell-cell adhesion protein found in normal and tumoral tissues. The aim of this study was to analyse Mel-CAM expression in mucoepidermoid carcinoma (MEC), and assess its importance in prognosis and its utility in differentiating high-grade MEC from squamous cell carcinoma (SCC). Immunohistochemical expression of Mel-CAM in 41 parotid MEC was correlated with clinical parameters. Ten cases of oral cavity SCC were included for comparison. Mel-CAM expression was found in 92.7% of the MEC but was not expressed by the SCC. Mel-CAM expression was greater in intermediate/high grade tumors, was weaker in patients that presented local recurrence, regional and distant metastasis, but no correlation between Mel-CAM and clinical stage and survival of the patients was found. Decreased Mel-CAM expression can impair cellular contact properties, facilitating growth, cell spreading and metastasis in MEC. Mel-CAM can also be useful in differentiating high grade MEC from SCC.

Asunto(s)

Antígenos CD , Carcinoma Mucoepidermoide/metabolismo , Glicoproteínas de Membrana/metabolismo , Proteínas de Neoplasias/metabolismo , Moléculas de Adhesión de Célula Nerviosa , Neoplasias de la Parótida/metabolismo , Adolescente , Adulto , Anciano , Biomarcadores de Tumor/metabolismo , Antígeno CD146 , Carcinoma Mucoepidermoide/diagnóstico , Carcinoma Mucoepidermoide/secundario , Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patología , Niño , Diagnóstico Diferencial , Femenino , Humanos , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Neoplasias de la Parótida/patología , Pronóstico

RESUMEN

BACKGROUND: Calcifying odontogenic cyst (COC) is an uncommon odontogenic lesion with few studies describing its immunohistochemical profile and proliferative activity reported in the literature. METHODS: Clinical and histological features and immunohistochemical expression of cytokeratins, Mel-CAM (CD146), bcl-2, PCNA and ki-67, in 10 cases of COC were studied. RESULTS: All 10 cases affected the maxilla, eight intraosseous and two peripheral. Five central cases were cystic and three were cystic associated with odontoma, and the two extraosseous showed solid histological pattern; immunohistochemistry was positive for cytokeratins 8, 14, 19, AE1/AE3 and 34betaE12 and bcl-2 in all cases, and Mel-CAM in six cases. Proliferative activity was greater in the epithelium of central cystic COC in relation to COC associated with odontoma and peripheral lesions. CONCLUSION: Calcifying odontogenic cysts showed odontogenic cytokeratin profile and bcl-2 and Mel-CAM expression indicate that these proteins may be involved in the development of COC. There were no recurrences after surgery, irrespective of their proliferative activity.

Asunto(s)

Antígenos CD , Neoplasias Maxilares/patología , Moléculas de Adhesión de Célula Nerviosa , Quiste Odontogénico Calcificado/patología , Antígenos de Diferenciación de Linfocitos T/análisis , Biomarcadores de Tumor/análisis , Antígeno CD146 , División Celular , Humanos , Inmunohistoquímica , Queratinas/análisis , Queratinas/clasificación , Antígeno Ki-67/análisis , Glicoproteínas de Membrana/análisis , Neoplasias Primarias Múltiples/patología , Odontoma/patología , Antígeno Nuclear de Célula en Proliferación/análisis , Proteínas Proto-Oncogénicas c-bcl-2/análisis