RESUMEN
More than 50% cells isolated from the endometrial cavity scraping and the myometrium of the rudimentary horn of an underdeveloped uterus removed from a patient with uterine aplasia and maintained under culturing conditions normal for mesenchymal stem cells (MSC) expressed embryonic transcription factors Oct4 and Nanog, embryonic cell membrane sialyl glycolipid SSEA4, and MSC markers. After 2-3 passages, the cells lost the expression of the early embryogenesis markers, but retained MSC markers. The presence of dormant stem cells in the underdeveloped endometrium and in the uterus indicates that this tissue has a regenerative potential that can be activated and used for completion of organ morphogenesis. This task requires the development of methods of early diagnosis of morphogenesis impairment and tools for safe reactivation of the ontogenesis.
Asunto(s)
Células Madre Embrionarias , Células Madre Mesenquimatosas , Útero , Femenino , Humanos , Diferenciación Celular/fisiología , Células Madre Embrionarias/metabolismo , Endometrio/metabolismo , Células Madre Mesenquimatosas/metabolismo , Miometrio , Útero/metabolismo , Útero/patologíaRESUMEN
The use of drug-loaded nanoparticles is an actively developed approach in targeted cancer therapy. Prevascularized spheroids generated from mesenchymal stem cells and endotheliocytes are considered as a model to evaluate the tropism of therapeutic nanoparticles to a specific tissue. Nanoparticles based on co-polymer of lactic and glycolic acids (poly(lactic-co-glycolic acid; PLGA) labeled with cyanine dye (Cy5) were incubated with prevascularized spheroids, and the rate of their penetration and their distribution in the spheroid-forming cells were evaluated. Endotheliocytes more intensively accumulated nanoparticles than mesenchymal stem cells: the number of nanoparticles in mixed-cell spheroids of mesenchymal stem cells and endotheliocytes was greater than in spheroids built solely of mesenchymal stem cells by 5±1.2 times. The developed 3D in vitro cell model provides a low-cost way to assess tissue tropism of therapeutic nanoparticles under conditions closer to natural in comparison with 2D culture.
Asunto(s)
Células Madre Mesenquimatosas , Nanopartículas , Copolímero de Ácido Poliláctico-Ácido Poliglicólico , Ácido Poliglicólico , Ácido Láctico/farmacología , Glicoles , Células CultivadasRESUMEN
Human placenta mesenchymal stromal cells were injected to healthy rats either stereotaxically into the striatum or intra-arterially through the internal carotid artery. Some cells injected into the brain migrated along the corpus callosum both medially and laterally or concentrated around small blood vessels. A small fraction of MSC injected intra-arterially adhered to the endothelium and stayed inside blood vessels for up to 48 hours mostly in the basin of the middle cerebral artery. Neither stereotaxic, nor intra-arterial transplantation of mesenchymal stromal cells modulated the proliferation of neural stem cells in the subventricular zone of the brain, but stereotaxic transplantation suppressed activation of their proliferation in response to traumatization with the needle.
Asunto(s)
Cuerpo Estriado/citología , Ventrículos Laterales/citología , Trasplante de Células Madre Mesenquimatosas/métodos , Células Madre Mesenquimatosas/citología , Células-Madre Neurales/citología , Placenta/citología , Animales , Arteria Carótida Interna/citología , Movimiento Celular , Proliferación Celular , Cuerpo Estriado/cirugía , Femenino , Humanos , Inyecciones Intraarteriales , Inyecciones Intraventriculares , Ventrículos Laterales/cirugía , Masculino , Células Madre Mesenquimatosas/fisiología , Arteria Cerebral Media/citología , Células-Madre Neurales/fisiología , Placenta/fisiología , Embarazo , Cultivo Primario de Células , Ratas , Ratas Wistar , Técnicas Estereotáxicas , Trasplante HeterólogoRESUMEN
We compared the effects of placental mesenchymal stromal cells and neural progenitor cells derived from induced human pluripotent cells after their intravenous administration to rats in 24 h after transitory occlusion of the middle cerebral artery. The therapeutic effects were evaluated by the dynamics of animal survival, body weight, neurological deficit, and the volume of infarction focus in 7, 14, 30, and 60 days after surgery. Intravenous injection of neural progenitor cells produced a therapeutic effect on the course of experimental ischemic stroke by increasing animal survival in the most acute period and accelerating compensation of neurological deficit and body weight recovery. Neural progenitor cells were more effective than mesenchymal stromal cells from human placenta. The effectiveness of intravenous transplantation of neural progenitor cells in the model of occlusion of the middle cerebral artery is shown by us for the first time, although the therapeutic effect of their direct transplantation into the brain has already been described.
Asunto(s)
Isquemia Encefálica/patología , Células Madre Mesenquimatosas/citología , Células-Madre Neurales/citología , Accidente Cerebrovascular/patología , Administración Intravenosa , Animales , Peso Corporal/fisiología , Células Cultivadas , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Trasplante de Células Madre Mesenquimatosas , Placenta , Embarazo , Ratas , Ratas WistarRESUMEN
The cells isolated from biopsy specimen of a patient with alcoholic liver cirrhosis and cultured under standard conditions for obtaining stromal cell culture clearly diverged during early passages into two morphologically and phenotypically different subtypes: epithelial and mesenchymal. Mesenchymal cells expressed CD90 and CD44 and epithelial cells expressed CD166, CD227, and hepatocyte growth factor receptor Met. Starting from passage 6, the culture underwent spontaneous morphological changes and by passages 8-10 contained only epithelium-like cells. CD90 and CD44 expression disappeared, CD166 and CD227 expression remained unchanged, and Met expression increased. A small fraction of cells expressed GATA-4, HNF3ß, HNF1α, and HNF4α. After addition of inducers of hepatogeneic differentiation, the cells started producing albumin.
Asunto(s)
Transición Epitelial-Mesenquimal/genética , Cirrosis Hepática Alcohólica/genética , Hígado/metabolismo , Células Madre Mesenquimatosas/metabolismo , Células Madre/metabolismo , Albúminas/biosíntesis , Albúminas/genética , Antígenos CD/genética , Antígenos CD/metabolismo , Biomarcadores/metabolismo , Diferenciación Celular , Proliferación Celular , Factor de Transcripción GATA4/genética , Factor de Transcripción GATA4/metabolismo , Expresión Génica , Factor Nuclear 1-alfa del Hepatocito/genética , Factor Nuclear 1-alfa del Hepatocito/metabolismo , Factor Nuclear 3-beta del Hepatocito/genética , Factor Nuclear 3-beta del Hepatocito/metabolismo , Factor Nuclear 4 del Hepatocito/genética , Factor Nuclear 4 del Hepatocito/metabolismo , Humanos , Hígado/patología , Cirrosis Hepática Alcohólica/metabolismo , Cirrosis Hepática Alcohólica/patología , Células Madre Mesenquimatosas/patología , Cultivo Primario de Células , Proteínas Proto-Oncogénicas c-met/genética , Proteínas Proto-Oncogénicas c-met/metabolismo , Células Madre/patologíaRESUMEN
We studied immunoregulatory properties of cultured human stem cells of mesenchymal and ectodermal origins after their administration to mice. Xenotransplantation of mesenchymal stem cells from human placenta reduced the number of CD11c(+)dendritic cells in mouse spleens, but did not affect activation of dendritic cells from mouse spleen in culture. It was also shown that splenocytes isolated from animals 10 days after transplantation of mesenchymal stem cells more actively proliferated in response to the polyclonal stimulation. At the same time, transplantation of neither mesenchymal nor neural stem cells affected the ratio of CD4(+)/CD8(+)T cells and their total content in the peripheral blood in comparison with the corresponding parameters in the control groups.
Asunto(s)
Ectodermo/citología , Mesodermo/citología , Trasplante de Células Madre , Células Madre/citología , Células Madre/inmunología , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Humanos , Inmunidad Celular/inmunología , Inmunohistoquímica , Masculino , Ratones , Ratones Endogámicos BALB CRESUMEN
Mesenchymal stem cells from human placenta obtained after term natural delivery were cultured and labeled with vital dye Dil of magnetic fluorescing microparticles. The labeled cells were transplanted intravenously to rats with occlusion of the median cerebral artery. Penetration of cells through the brain-blood barrier and their distribution in the brain of experimental animals were studied on serial cryostat sections. Two models of cerebral artery occlusion associated with different traumatic consequences were used. The efficiency of crossing the blood-brain barrier by transplanted cells, the number of mesenchymal cells attaining the ischemic focus and neurogenic zones, and the time of death of transplanted cells largely depended on the degree and nature of injury to the central nervous system, which should be taken into account when planning the experiments for evaluation of the effects of cell therapy on the models of neurological diseases and in clinical studies in the field of regenerative neurology.
Asunto(s)
Isquemia Encefálica/terapia , Sistema Nervioso Central/citología , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/fisiología , Accidente Cerebrovascular/terapia , Animales , Barrera Hematoencefálica/fisiología , Diferenciación Celular , Modelos Animales de Enfermedad , Femenino , Humanos , Nanopartículas de Magnetita , Placenta/citología , Embarazo , Ratas , Trasplante HeterólogoRESUMEN
Mesenchymal stem cells enzymatically isolated from human placenta were labeled with magnetic fluorescent microparticles (d=0.96 µ). We showed that microparticles in high doses (>10 µl stock suspension per 1 ml culture medium) significantly inhibited cell proliferation in culture. In our work we determined the optimal concentration of particles not affecting physiological properties of mesenchymal stem cells: it does not change cell proliferation, does not induce apoptosis, and does not modulate their transdifferentiation into neuronal cells. In vivo experiments showed that the chosen particles allow easy visualization of transplanted cells ex vivo on sections of different tissues.
Asunto(s)
Magnetismo , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de los fármacos , Nanopartículas/efectos adversos , Animales , Apoptosis/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Femenino , Humanos , Masculino , Nanopartículas/química , Embarazo , Ratas , Ratas WistarRESUMEN
Cultures of mesenchymal cells from human decidual tooth pulp were derived. The phenotype and capacity to osteogenic and adipogenic differentiation of these cells are close to those of bone marrow mesenchymal stem cells. Decidual tooth pulp mesenchymal cells populate biodegraded polylactide scaffolds and hence, can be used for the creation of tissue engineering transplants for bone defect repair. Storage of decidual tooth pulp mesenchymal cells in the stem cell cryobanks together with umbilical blood will appreciably extent the periods of age for collection of juvenile autologous stem cells for use throughout the life span.
Asunto(s)
Huesos , Pulpa Dental/citología , Células Madre Mesenquimatosas/citología , Ingeniería de Tejidos/métodos , Diente Primario/citología , Técnicas de Cultivo de Célula/métodos , Diferenciación Celular/fisiología , Citometría de Flujo , Humanos , Inmunohistoquímica , Microscopía Fluorescente , Poliésteres , Andamios del TejidoRESUMEN
It is demonstrated that the output optical signal of MTT test is directly proportional to the number of viable cells in the primary culture of mesenchymal cells (skin fibroblasts and mesenchymal stem cells from the bone marrow, placenta, and umbilical cord). The slope of the best curve in coordinates "cell number - optical signal" reflecting specific productivity of MTT-formazan characterizes mean dehydrogenase activity of cells and their physiological activity. It was found that in vitro dehydrogenase activity of primary cultures of mesenchymal cells increased during the first 3-5 passages and then tended to decrease. The variant of MTT method presented here can be used for standardization of cell materials.
Asunto(s)
Técnicas de Cultivo de Célula/normas , Oxidorreductasas/metabolismo , Células de la Médula Ósea/fisiología , Células Cultivadas , Femenino , Formazáns/metabolismo , Humanos , Células Madre Mesenquimatosas/fisiología , Embarazo , Sales de Tetrazolio/metabolismo , Tiazoles/metabolismoRESUMEN
A method for preparation and expansion of mixed keratinocyte and melanocyte culture from human skin biopsy specimens was developed. The culture contains two melanocyte types: derivatives of hair follicle stem cells and epidermal basal layer stem cells. Fibroblasts were completely eliminated after culturing in selective medium.
Asunto(s)
Queratinocitos/citología , Melanocitos/citología , Pigmentación de la Piel , Vitíligo/patología , Biopsia , Técnicas de Cocultivo , HumanosRESUMEN
Mesenchymal stem cells isolated from human placenta and in vitro labeled with fluorescent magnetic microparticles were intravenously injected to rats 2 days after induction of focal cerebral ischemia (endovascular model). According to MRT findings, transplantation of mesenchymal stem cells led to an appreciable reduction of the volume of ischemic focus in the brain. Two or three weeks after transplantation, labeled cells accumulated near and inside the ischemic focus, in the hippocampus, and in the subventricular zone of both hemispheres. Only few human mesenchymal stem cells populating the zone adjacent to the ischemic focus started expressing astroglial and neuronal markers. On the other hand, transplantation of mesenchymal stem cells stimulated proliferation of stem and progenitor cells in the subventricular zone and migration of these cells into the ischemic zone. Positive effects of transplantation of these cells to rats with experimental ischemic stroke are presumably explained by stimulation of proliferation of resident stem and progenitor cells of animal brain and their migration into the ischemic tissue and adjacent areas. Replacement of damaged rat neurons and glial cells by transplanted human cells, if it does take place, is quite negligible.
Asunto(s)
Isquemia Encefálica/terapia , Células Madre Mesenquimatosas/citología , Placenta/citología , Accidente Cerebrovascular/terapia , Animales , Diferenciación Celular/fisiología , Proliferación Celular , Células Cultivadas , Modelos Animales de Enfermedad , Femenino , Humanos , Masculino , Trasplante de Células Madre Mesenquimatosas , Células Madre Mesenquimatosas/fisiología , Embarazo , RatasRESUMEN
The results of the development and introduction of universal production standards for cell products of mesenchymal origin are presented: technology for obtaining and culturing of primary cell cultures from human postnatal organs and tissues, cell product quality and safety control procedures, methods for cell product storage and transportation, and the necessary files.
Asunto(s)
Productos Biológicos/aislamiento & purificación , Productos Biológicos/normas , Ingeniería Biomédica/normas , Células Madre Mesenquimatosas/citología , Productos Biológicos/biosíntesis , Células de la Médula Ósea/citología , Técnicas de Cultivo de Célula , Separación Celular/métodos , Separación Celular/normas , Estructuras Celulares , Sangre Fetal/citología , Fibroblastos/citología , Humanos , Control de Infecciones/métodos , Control de Calidad , Piel/citología , Manejo de Especímenes/métodos , Manejo de Especímenes/normas , TransportesRESUMEN
Cell-mediated cytotoxicity was studied in vitro during the interaction of bone marrow mesenchymal stem cells, fibroblast-like cells from newborn umbilical cord, and skin fibroblasts of an adult donor with peripheral blood mononuclear cells. Independently on the origin, mesenchymal cells were not lysed with allogeneic natural killer cells and cytotoxic lymphocytes. Mixed cultures of mesenchymal cells had no cytotoxic effect on peripheral blood mononuclear cells and did not activate proliferation of T and B lymphocytes, natural killer cells, and CD14+ lymphocytes. In vitro experiments showed that mesenchymal cells of different origin and allogeneic immunocompetent cells are tolerant to each other.
Asunto(s)
Células Madre Mesenquimatosas , Adulto , Células de la Médula Ósea/citología , Células de la Médula Ósea/inmunología , Células Cultivadas , Fibroblastos/citología , Fibroblastos/inmunología , Antígenos HLA/inmunología , Humanos , Leucocitos Mononucleares/citología , Leucocitos Mononucleares/inmunología , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/inmunología , Piel/citología , Cordón Umbilical/citologíaRESUMEN
We compared the capacity of cultured human skin fibroblasts, human umbilical cord cells obtained after normal delivery on gestation week 38-40, and mesenchymal bone marrow stem cells to differentiation into adipocytes, osteoblasts, and chondrocytes. Our findings suggest that mesenchymal stem cells are multipotent cells and can differentiate into adipose, cartilaginous, and bone tissue. Umbilical cord fibroblast-like cells can differentiate into adipocytes and chondrocytes, and only few cells in this culture can differentiate into osteoblasts. Skin fibroblasts differentiate only into adipocytes.
Asunto(s)
Células de la Médula Ósea/citología , Fibroblastos/citología , Células Madre Mesenquimatosas/citología , Mesodermo/citología , Piel/citología , Cordón Umbilical/citología , Adipocitos/citología , Células de la Médula Ósea/metabolismo , Diferenciación Celular , Células Cultivadas , Condrocitos/citología , Femenino , Fibroblastos/metabolismo , Citometría de Flujo , Humanos , Inmunohistoquímica , Células Madre Mesenquimatosas/metabolismo , Células Madre Multipotentes/citología , Células Madre Multipotentes/metabolismo , Osteoblastos/citologíaRESUMEN
The expression of cytoplasmic and surface proteins in cultured human skin fibroblasts, human umbilical cord cells obtained after normal delivery on gestation week 38-40, and mesenchymal bone marrow stem cells was compared by the methods of immunocytochemistry and flow cytofluorometry. Bone marrow mesenchymal stem cells expressed a great variety of marker proteins typical of stem and progenitor cells and did not express proteins typical of differentiated cells. Fibroblast-like umbilical cord cells expressed markers of both stem cells and differentiated cells. Fibroblasts of dermal origin were characterized by intensive expression of proteins typical of differentiated cells.
Asunto(s)
Mesodermo/citología , Células Madre/citología , Cordón Umbilical/citología , Células de la Médula Ósea/citología , Células de la Médula Ósea/metabolismo , Linaje de la Célula , Femenino , Fibroblastos/citología , Fibroblastos/metabolismo , Humanos , Inmunohistoquímica , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/metabolismo , Proteoma/biosíntesis , Piel/citología , Células Madre/metabolismoRESUMEN
Expression of markers, collagens, and HLA-1 by human skin fibroblasts and fibroblast-like cells isolated from the umbilical Wharton's jelly was compared. Skin fibroblasts express collagens (proteins characteristic of differentiated cells of this histogenetic series) and HLA-1, while umbilical cells express, in addition to collagens, juvenile surface markers and almost no HLA-1. This indicates that fibroblast-like cells isolated from different sources are different and can serve as sources for the creation of cell preparations with different characteristics in future.
Asunto(s)
Colágeno Tipo II/análisis , Colágeno Tipo I/análisis , Fibroblastos/química , Antígeno HLA-A1/análisis , Piel/citología , Cordón Umbilical/citología , Antígeno AC133 , Animales , Antígenos/análisis , Antígenos CD/análisis , Antígenos CD34/análisis , Biomarcadores/análisis , Femenino , Fibroblastos/trasplante , Glicoproteínas/análisis , Humanos , Inmunohistoquímica , Proteínas de Filamentos Intermediarios/análisis , Proteínas del Tejido Nervioso/análisis , Nestina , Péptidos/análisis , Ratas , Ratas Wistar , Vimentina/análisis , Factor de von Willebrand/inmunologíaRESUMEN
Comparative analysis of the expression of some surface markers of human bone marrow mesenchymal stem cells, umbilical fibroblast-like cells, and skin fibroblasts was carried out by the flow cytofluorometry method. Mesenchymal stem cells and umbilical fibroblast-like cells were similar by the levels of expression of the main histocompatibility complex antigens, adhesion molecules, and some growth factor receptors. The profile of skin fibroblast surface antigens was characterized by higher expression of the markers typical of differentiated cells. The results prove the possibility of using umbilical fibroblast-like cells as an alternative source of mesenchymal stem cells for cell replacement therapy.
Asunto(s)
Células de la Médula Ósea/citología , Fibroblastos/citología , Células Madre Mesenquimatosas/citología , Antígenos CD/análisis , Células de la Médula Ósea/inmunología , Sangre Fetal/citología , Fibroblastos/inmunología , Citometría de Flujo/métodos , Antígenos HLA/análisis , Humanos , Inmunohistoquímica , Células Madre Mesenquimatosas/inmunología , Fenotipo , Piel/citología , Piel/inmunologíaRESUMEN
The paper summarizes the results of HIV-1 serotyping by using 56 positive sera collected in the territory of the Republic of Tajikistan (RT) from intravenous drug users (IVDU). It was made by solid-phase ELISA based on synthetic peptides, mimicking different variants of the apical epitope of the HIV-1 gp120 V3-loop. Two types of conjugates, those specific to human IgG and IgA, were used to detect the immune complexes. Serotypes, as determined according to IgG and IgA-based ELISA, coincided, however, the latter were proven to be more suitable for serotyping. There is a high level of HIV-1 serotype heterogeneity among IVDU in RT; altogether, 4 serotypes were identified, i.e. B (10%), B+A/C (18%), A/C (20%) and A/C+B (52%). The modern serotype HIV-1 diversity in RT resembles the epidemiologic situation in the territory of the former USSR as observed in the late 80-ies-early 90-ies of the last century.