RESUMEN
During open heart procedures, the heart is stopped. The surgeon needs a blood-free, motionless work area. The heart-lung bypass machine provides circulation for the rest of the body during surgery. The perfusionist, as an active member of the surgical team, operates the heart-lung bypass machine. Historical development of the perfusionist's role and collaboration with other members of the healthcare team are discussed.
Asunto(s)
Puente Cardiopulmonar , Perfusión , Humanos , Perfusión/enfermeríaRESUMEN
BACKGROUND: This investigation was undertaken to establish a serum-free organ culture technique allowing for the morphological and physiological maintenance of human fetal stomach in vitro. METHODS: Explants from gastric corpuses (12-17 weeks of gestation) were cultured in serum-free medium for periods of up to 15 days. RESULTS: After 15 days of culture, surface mucous cells were more mature, gastric glands were numerous and well developed, and all epithelial cell types were morphologically very well preserved. Morphometric measurements of the glands revealed an accelerated development in culture compared with that found in utero. Even though the incorporation of [3H]thymidine into total DNA decreased, the labeling indices determined by radioautography confirmed that epithelial cell proliferation was maintained especially in the pit/neck portion and at the base of the glandular compartments. A significant increase in total glycoprotein synthesis, as evaluated by the incorporation of [3H]glucosamine, was observed and correlated with the differentiation of the mucous cells. CONCLUSIONS: This investigation establishes for the first time that human gastric mucosa can be maintained up to 15 days in organ culture and that maturation of the gastric mucosa can be reproduced in chemically defined media.
Asunto(s)
Mucosa Gástrica/embriología , Diferenciación Celular , ADN/biosíntesis , Femenino , Madurez de los Órganos Fetales , Mucosa Gástrica/citología , Mucosa Gástrica/ultraestructura , Glicoproteínas/biosíntesis , Humanos , Técnicas de Cultivo de Órganos , Embarazo , Biosíntesis de ProteínasRESUMEN
The purpose of tumour staging for colorectal cancer (CRC) is to help define clinical management, facilitate communication between physicians, provide a basis for stratification and analysis of treatment results in prospective studies, and provide some prognostic information for patients and their families. The World Congresses of Gastroenterology, Digestive Endoscopy, and Coloproctology, Working Party on staging for CRC studied six commonly used systems to review their strengths and weaknesses. Although it was concluded that defining a new staging system was unnecessary, it was recognized that there is a need to define a terminology to describe the full anatomic extent of spread of CRC. Furthermore, we note that there are several additional features, derived from both clinical and pathology information, which have had prognostic significance shown by appropriately constructed multivariate analyses and which can be used to formulate a more accurate prognostic index than that provided by a description of anatomical tumour spread. Thus the Working Party came to two principal conclusions. First, a standard format should be adopted for the collection of the essential data required for prospective studies, and we recommend the 'International Documentation System (IDS) for CRC' for this purpose. Second, a nomenclature which describes the full anatomical extent of tumour spread and residual tumour status in CRC has been defined and should be adopted, from which all currently used staging systems can be derived. We have called this nomenclature the 'International Comprehensive Anatomical Terminology (ICAT) for CRC'. In the event that these recommendations are adopted, we envision that there will be improved clarity in the documentation of treatment outcome for patients with CRC and improved communication of results derived from prospective studies. Furthermore, an acceptance of IDS and ICAT would set the scene to develop a prognostic index for individual patients with CRC by the expansion of anatomical clinicopathology staging information to include additional factors which have independent prognostic significance.
Asunto(s)
Neoplasias Colorrectales/patología , Recolección de Datos/métodos , Terminología como Asunto , Humanos , Estadificación de Neoplasias/métodos , Pronóstico , Estadística como AsuntoRESUMEN
Accidental hypothermia due to exposure is an infrequent cause of circulatory arrest. A premature diagnosis of clinical death must be avoided in these patients and vigorous attempts at active rewarming are indicated. Extracorporeal circulation in the form of partial cardiopulmonary bypass has been reported as an effective means of rapid, even core rewarming. We wish to report a recent case at Ruby Memorial Hospital in which extracorporeal circulation was used successfully in resuscitating a profoundly hypothermic multi-trauma victim.
Asunto(s)
Accidentes de Tránsito , Circulación Extracorporea , Hipotermia/complicaciones , Traumatismo Múltiple/complicaciones , Resucitación/métodos , Adulto , Femenino , HumanosRESUMEN
The influence of epidermal growth factor (EGF) and hydrocortisone on the functional development of human fetal colon was studied in organ cultures. Fetal colon (14 to 17 weeks gestation) was cultured for 5 days at 37 degrees C in serum-free Leibovitz L-15 medium alone or supplemented with 1, 10, and 100 ng of EGF/ml or with 50 ng of hydrocortisone/ml of culture medium. The overall morphology of the colonic explants was not altered by the hormonal addition. In the continuous presence of EGF (1, 10, and 100 ng/ml) for 5 days, a significant decrease of [3H]thymidine incorporation into DNA was observed. At the brush border level, the addition of EGF induced a significant drop in sucrase, maltase, and alkaline phosphatase activities. These enzymic modifications occurred between the third and fifth day of culture, whereas variation in DNA synthesis was already evident within 24 h. The addition of hydrocortisone at a dose affecting the small intestine (50 ng/ml) did not significantly influence colonic DNA synthesis nor the digestive enzymic activities. These observations show for the first time that EGF, but not hydrocortisone, influences the proliferation and differentiation of human fetal colonic mucosa.
Asunto(s)
Colon/embriología , Factor de Crecimiento Epidérmico/farmacología , Hidrocortisona/farmacología , División Celular/efectos de los fármacos , Colon/efectos de los fármacos , Colon/enzimología , Medios de Cultivo , Feto , Humanos , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/embriología , Mucosa Intestinal/ultraestructura , Microscopía Electrónica de Rastreo , Técnicas de Cultivo de ÓrganosRESUMEN
Cell proliferation during morphogenesis of human stomach was investigated using radioautography and biochemical determinations of [3H]-thymidine incorporation into DNA. Labeling indices in the epithelium, mesenchyme and muscle layer were established on radioautographs and the heights (mm) of the gastric glands were measured between 10 and 17 weeks of gestation. At 11-12 weeks, the appearance of the first pit/gland was noted, and the labeling index ranged from 9.2 to 10.2%. Labeled cells were present at all levels of the stratified epithelium. Between 14 and 16 weeks, the total epithelial labeling index declined sharply (8.1 to 5.4%) with a concomitant increase of the height of the pit/gland structures (0.055 to 0.080 mm). High proliferative activity was also recorded in the mesenchyme and the muscle layer, the labeling indices decreasing between 10 and 17 weeks. The biochemical quantitation of the [3H]-thymidine uptake into the total gastric DNA clearly supported the continuous decrease of the cell proliferation determined by radioautography. Detailed analysis of the epithelium showed that proliferative cells were more numerous at the base of the gland at the earliest stage (11 weeks) but concentrated in the pit/neck regions by 13-14 weeks. As the pit/gland development proceeded (14 to 17 weeks) labeled cells remained more abundant in the pit/neck regions of the gland (10.9%) and were rarely seen on the surface epithelium (2%). The present investigation provides basic quantitative data regarding cell proliferation in developing human stomach, and indicates that the morphogenesis of the gastric glands is correlated with the high proliferative capacity of the pit/neck cells.
Asunto(s)
Estómago/citología , División Celular , ADN/biosíntesis , Células Epiteliales , Epitelio/embriología , Epitelio/ultraestructura , Feto/citología , Feto/metabolismo , Mucosa Gástrica/citología , Mucosa Gástrica/embriología , Mucosa Gástrica/ultraestructura , Humanos , Mesodermo/citología , Mesodermo/metabolismo , Mesodermo/ultraestructura , Microscopía Electrónica , Morfogénesis , Estómago/embriología , Estómago/ultraestructura , Timidina/metabolismo , TritioRESUMEN
Cell proliferation during morphogenesis of human colon from 8 to 18 weeks of gestation was investigated using radioautography. The sites of [3H]-thymidine uptake were localized in the epithelium, the mesenchyme, and the muscularis externa. As long as the epithelium was stratified, the proliferative pool was very large, and the highest labeling indices (up to 28%) were found during this period. With the formation of villi, the proliferative pool decreased and focused exclusively in the developing crypts by 14-15 weeks. Between 8 and 11 weeks, the labeling index was similar both in the epithelium and the mesenchyme. By 13 weeks of gestation, the small intestinal cell proliferation pattern (epithelium greater than mesenchyme greater than muscular layer) was established in the fetal colon. The biochemical quantitation of the [3H]-thymidine uptake into the total colonic DNA supported the decreasing cell proliferation pattern determined by radioautography. The present investigation establishes basic quantitative data regarding cell proliferation during a particular developmental phase of the human colon.
Asunto(s)
Colon/embriología , Autorradiografía , Diferenciación Celular , División Celular , Colon/citología , Técnicas de Cultivo , ADN/biosíntesis , Feto , Humanos , MorfogénesisRESUMEN
Cell proliferation in the developing human esophagus was investigated by autoradiography using organ culture. The sites of [3H]-thymidine uptake were localized in the epithelium, the mesenchyme, and the muscularis externa of fetal esophageal explants from 10 to 16 weeks of gestation. Proliferating cells were abundant throughout the stratified epithelium at 10 weeks of gestation. Many labeled nuclei in the mesenchyme and the muscular layer were observed. With the development of the stratified columnar ciliated epithelium, a confinement of the proliferating zone in the basal cell layers occurred, and ciliated cells never appeared labeled. The quantitation of proliferating cells showed a labeling index at its highest value between 10 and 12 weeks; this drastically decreased between 12 and 14 weeks' gestation. A similar pattern was noted for the mesenchyme, while the labeling index in the muscularis externa peaked during the 11-14 weeks period. In all fetuses, the highest labeling index was always recorded in the epithelium. The biochemical quantitation of the [3H]-thymidine uptake into the total esophageal DNA clearly supported the continuous decrease of the cell proliferation determined by autoradiography between 10 and 16 weeks of gestation. The present investigation provides for the first time basic quantitative data regarding cell proliferation during a particular developmental phase of the human esophagus.
Asunto(s)
Esófago/embriología , División Celular , Replicación del ADN , Edad Gestacional , Humanos , Técnicas de Cultivo de Órganos , Timidina/análisisRESUMEN
The influence of epidermal growth factor (EGF) on the differentiation and proliferation of human fetal jejunum was studied in organ cultures. Fetal intestine (11-14-wk gestation) was cultured for 5 days at 37 degrees C in serum-free Leibovitz L-15 medium alone or supplemented with 25, 50, and 100 ng EGF/ml culture medium. The addition of hormone did not modify the morphology of the intestinal explants. Biochemical studies revealed that lactase activity was significantly increased with the addition of 50 and 100 ng EGF/ml culture medium. On the other hand, the increase in sucrase, trehalase, and glucoamylase activities that normally occurs during the culture was repressed in the presence of increasing concentrations of EGF. Deoxyribonucleic acid synthesis was significantly decreased after 5 days of culture even in the presence of the lowest EGF concentration used. Concomitantly, the labeling index of the epithelial cells dropped drastically in the presence of EGF. The EGF-induced variation in DNA synthesis was already evident within 24 h of culture, whereas enzymic modifications occurred only between the third and fifth day of culture. The simultaneous addition of EGF and hydrocortisone (50 ng/ml) did not reveal any synergistic action between the two hormones on the hydrolytic activities of the brush border. However, EGF did inhibit hydrocortisone-stimulated DNA synthesis. The present work provides for the first time some basic data on the influence of EGF on brush border hydrolytic activities and on epithelial cell proliferation of human fetal jejunum. These observations strongly suggest that EGF plays an important role in the fetal development of the human gastrointestinal tract.
Asunto(s)
División Celular/efectos de los fármacos , ADN/efectos de los fármacos , Factor de Crecimiento Epidérmico/farmacología , Feto/efectos de los fármacos , Yeyuno/efectos de los fármacos , Células Cultivadas , ADN/biosíntesis , Sinergismo Farmacológico , Feto/metabolismo , Humanos , Hidrocortisona/farmacología , Hidrólisis , Yeyuno/metabolismoRESUMEN
The purpose of this work was to study the human fetal esophagus maintained in organ culture. Esophageal explants from 8 fetuses aged from 12 to 16 weeks of gestation were cultured up to 21 days at 37 degrees C in Leibovitz L-15 serum-free medium. Between 12 and 16 weeks of gestation, the esophagus has a stratified columnar ciliated epithelium, and glycogen aggregates are present in all cell layers. This morphology remains the same up to 5 days in culture. After 7 to 9 days, a vacuolization in the upper half layer occurs, leading to a lifting off of the ciliated layer and a flattening of the subjacent cells. After 15 days of culture, the esophageal epithelium is stratified squamous and the cells are exfoliated at the surface of the explants. Glycogen aggregates are still present in all layers. Islets of ciliated cells resting on the basal cell layers develop within the squamous epithelium. With the extension of the culture period up to 21 days, the general morphology of the epithelium does not change. The ultrastructural features of the newly formed squamous epithelium, with its basal lamina, are similar to that reported for human adult esophageal epithelium. During the course of the culture, the DNA synthesis continues as determined by autoradiography. It is concluded that it is possible to maintain viable human fetal esophagus in organ culture and that an accelerated maturation takes place leading to the formation of the adult esophageal epithelium.
Asunto(s)
Esófago/embriología , Diferenciación Celular , Esófago/ultraestructura , Edad Gestacional , Humanos , Técnicas de Cultivo de ÓrganosRESUMEN
The effects of epidermal growth factor (EGF), cortisone and thyroxine on deoxyribonucleic acid (DNA) synthesis in the esophagus, stomach, small intestine and colon have been studied in suckling mouse. Daily administration of EGF [4 micrograms/g body weight (bw)/day] during 3 days to 8-day-old mice induced a significant increase of the incorporation of [3H]thymidine into DNA in the stomach, the small intestine, and the two halves of the colon. The DNA synthesis in the esophagus remained unaffected by the EGF treatment. The maximal increase of [3H]thymidine incorporation into DNA was observed in the colon, and represented 112%. Daily administration of cortisone acetate (25 micrograms/g bw/day) or thyroxine (1 microgram/g bw/day) during 3 days to 8-day-old mice had no significant influence of the DNA synthesis of any part of the gastrointestinal tract. These results show that EGF is able to affect the DNA synthesis in the stomach, small intestine and colon of suckling mice.
Asunto(s)
ADN/biosíntesis , Sistema Digestivo/efectos de los fármacos , Factor de Crecimiento Epidérmico/farmacología , Animales , Cortisona/farmacología , Sistema Digestivo/metabolismo , Ratones , Ratones Endogámicos ICR , Tiroxina/farmacología , Distribución TisularRESUMEN
Cell proliferation of the developing human jejunum was investigated by radioautography using organ culture. The sites of [3H]-thymidine uptake were localized in the epithelium, the mesenchyme and the muscularis externa of fetal human small intestinal explants from 8 to 18 weeks of gestation. Proliferating cells were abundant and scattered throughout the stratified epithelium before the appearance of villi. Many nuclei in the mesenchyme and the muscular layer were labeled. With the villi formation and the simplification of the epithelium, there was a confinement of the proliferating zone in the intervillus areas and developing crypts. The quantitation of proliferating cells showed a labeling index at its highest value between 8 and 10 weeks of gestation decreasing gradually up to 18 weeks of gestation at the epithelial and mesenchymal level. In the muscularis externa, the labeling remained more or less constant between 11 and 18 weeks of gestation. The organ culture of intestinal explants for 5 days did not modify epithelial cell proliferation. The present investigation provides for the first time basic quantitative data regarding cell proliferation in the developing human jejunum.
Asunto(s)
Yeyuno/citología , Autorradiografía , División Celular , Feto/citología , Edad Gestacional , Humanos , Yeyuno/embriología , Yeyuno/metabolismo , Técnicas de Cultivo de Órganos , Timidina/metabolismoRESUMEN
Human fetal colon (14-16 weeks gestation) was cultured as explants for 15 days in serum-free Leibovitz L-15 medium at 37 degrees C. The overall morphology of the colonic explants was well maintained throughout the culture period and all epithelial cell types retained their ultrastructural characteristics. The incorporation of [3H]-leucine continued and even increased, reflecting sustained synthesis of proteins. Even though the incorporation of [3H]-thymidine into the total DNA decreased during culture, the synthesis of DNA continued. The sites of [3H]-thymidine incorporation into the different layers of the colonic wall were studied by radioautography. The incorporation of the radioactive precursor occurs mainly in the epithelium and to lesser degrees in the mesenchyme and the muscular layer. Labeled epithelial nuclei were located in the intervillous areas but not on the villi. The labeling index of the epithelial cells remained constant throughout the culture period indicating the preservation of the proliferative capacity of the epithelium. Brush-border hydrolytic activities, namely those of sucrase, maltase, lactase, trehalase, glucoamylase and alkaline phosphatase, were assayed in the colonic tissue. These enzymic activities generally decreased in the tissue and increased in the medium during the course of culture. These observations clearly demonstrate that fetal colon can be maintained viable for at least 15 days in a serum-free medium. Organ culture now provides the opportunity to study the normal function and metabolism of human colon during its development.
Asunto(s)
Colon/embriología , Autorradiografía , Colon/metabolismo , ADN/biosíntesis , Humanos , Hidrólisis , Microvellosidades/metabolismo , Técnicas de Cultivo de Órganos , Biosíntesis de ProteínasRESUMEN
The direct influence of epidermal growth factor (EGF) on the differentiation and proliferation of small intestine was studied in organ culture. Eight-day-old mouse small intestine was cultured during 2 days in serum-free Leibovitz L-15 medium alone or supplemented with EGF (50, 100, and 500 ng/ml) either at room temperature or at 37 degrees C. Brush border membrane hydrolytic activities, namely, sucrase, lactase, glucoamylase, trehalase, maltase, and alkaline phosphatase, were assayed in the intestinal tissue as well as in the culture medium. None of the brush border enzymic activities was affected by the addition of EGF to the culture medium. This lack of effect is not temperature dependent since it occurred both at room temperature and at 37 degrees C. The addition of hydrocortisone (10(-6) M) to the culture medium induced the appearance of sucrase activity and increased the activity of the other brush border enzymes. The simultaneous addition of EGF with hydrocortisone did not influence the response of the intestinal explants to hydrocortisone. The deoxyribonucleic acid (DNA) content was determined while DNA synthesis was evaluated by the incorporation of (3H)-thymidine. The addition of EGF did not affect DNA content or (3H)-thymidine incorporation into DNA either at room temperature or at 37 degrees C. The EGF binding to epithelial cells did not significantly vary throughout the culture period and a down-regulation process occurred in presence of EGF. These observations strongly suggest that EGF does not act as a primary cue for inducing developmental changes in suckling mouse small intestine. It is proposed that EGF induces a systemic reaction in vivo that then influences the neonatal small intestine.
Asunto(s)
Factor de Crecimiento Epidérmico/farmacología , Yeyuno/efectos de los fármacos , Animales , Animales Lactantes , ADN/efectos de los fármacos , Factor de Crecimiento Epidérmico/metabolismo , Hidrocortisona/farmacología , Yeyuno/crecimiento & desarrollo , Yeyuno/metabolismo , Ratones , Ratones Endogámicos ICR , Microvellosidades/efectos de los fármacos , Microvellosidades/enzimología , Técnicas de Cultivo de ÓrganosRESUMEN
Vaginal discharges in prepubertal girls can be categorized under two broad headings--those with specific microbiological causes and, in the absence of such, those that are nonspecific in origin. For specific vulvovaginitis, treatment should be tailored to the findings on cultures, wet mounts, KOH, or other slide preparations. For the sexually transmissible organisms resulting in a vaginal discharge, thorough social service investigation should be undertaken in addition to appropriate antibiotic therapy. When a microbiological cause cannot be found and a foreign body has been ruled out, one is left with a diagnosis of nonspecific vulvovaginitis; treatment goals should be aimed at reassuring and re-educating the patient and parents in good hygienic practices as well as the elimination of potential irritants.
Asunto(s)
Vulvovaginitis/etiología , Adolescente , Candidiasis/diagnóstico , Candidiasis/patología , Candidiasis/terapia , Niño , Infecciones por Chlamydia/tratamiento farmacológico , Infecciones por Chlamydia/patología , Femenino , Gonorrea/tratamiento farmacológico , Gonorrea/patología , Infecciones por Haemophilus/tratamiento farmacológico , Humanos , Recién Nacido , Leucorrea/etiología , Leucorrea/microbiología , Oxiuriasis/diagnóstico , Oxiuriasis/tratamiento farmacológico , Tricomoniasis/tratamiento farmacológico , Tricomoniasis/patología , Vagina/microbiología , Vulvovaginitis/diagnóstico , Vulvovaginitis/tratamiento farmacológico , Vulvovaginitis/microbiologíaRESUMEN
The influence of hydrocortisone on the differentiation and proliferation of human fetal small intestine was studied. Fetal intestine (12- to 14-week gestation) was cultured during 5 days at 37 degrees C in serum-free Leibovitz L-15 medium alone or supplemented with hydrocortisone (12.5, 25, and 50 ng/ml). The addition of different concentrations of hormone did not affect the morphology of the intestinal explants. Brush border membrane hydrolytic activities, namely, sucrase, lactase, glucoamylase, trehalase, and alkaline phosphatase activities, were assayed in the intestinal tissue. A specific increase of lactase and alkaline phosphatase activities was induced by the addition of 25 and 50 ng hydrocortisone/ml culture medium. The DNA synthesis evaluated by the incorporation of [3H]thymidine was increased by the addition of 50 ng hydrocortisone/ml. The sites of incorporation into the different layers of the intestinal wall were studied by radioautography. The incorporation of the radioactive precursor occurred mainly in the epithelium and to a lesser degree in the mesenchyme and muscular layers. Labeled epithelial nuclei were located in the intervillous areas and developing crypts but not on the villi. The addition of hydrocortisone induced a significant increase of the labeling index of the epithelial cells. The present work provided for the first time some basic data on the influence of hydrocortisone on brush border hydrolytic activities and on epithelial cell proliferation of human fetal small intestine.
Asunto(s)
Hidrocortisona/farmacología , Intestino Delgado/efectos de los fármacos , Diferenciación Celular/efectos de los fármacos , División Celular/efectos de los fármacos , ADN/biosíntesis , Disacaridasas/metabolismo , Células Epiteliales , Epitelio/efectos de los fármacos , Epitelio/metabolismo , Feto , Edad Gestacional , Humanos , Intestino Delgado/citología , Intestino Delgado/metabolismo , Microvellosidades/metabolismo , Técnicas de Cultivo de ÓrganosRESUMEN
The postnatal development of enteropeptidase activity has been examined on mucosal scrapping of the proximal part of the mouse small intestine. The activity was present at birth and remained low during the first 15 days of life. Then it rapidly increased reaching adult level within 2 days. Daily administration of cortisone acetate (25 micrograms X g body weight (bw)-1 X day-1), insulin (12.5 mU X g bw-1 X day-1), or epidermal growth factor (4 micrograms X g bw-1 X day-1) during 3 days to 8-day-old mice induced a premature increase of enteropeptidase. The maximal increase was observed with cortisone treatment, the enzymic activity representing 70% of the adult level. Thyroxine alone (1 microgram X g bw-1 X day-1) had no significant effect on enteropeptidase activity. Hormonal interactions have been evaluated by studying the effects of different hormonal combinations. Finally, cortisone acetate which has a major effect on this activity during suckling period was unable to influence adult small intestinal enteropeptidase activity.
Asunto(s)
Endopeptidasas/metabolismo , Enteropeptidasa/metabolismo , Hormonas/farmacología , Intestino Delgado/enzimología , Animales , Animales Lactantes , Cortisona/farmacología , Factor de Crecimiento Epidérmico/farmacología , Insulina/farmacología , Intestino Delgado/crecimiento & desarrollo , Ratones , Ratones Endogámicos ICR , Tiroxina/farmacologíaRESUMEN
Human fetal intestine (10-14 wk gestation) has been cultured as explants in a serum-free Leibovitz L-15 medium for periods up to 9 days. As determined by light microscopy, the overall architecture of the intestinal explant was maintained throughout the culture period. At the ultrastructural level the villus absorptive cells remained tall with well-defined brush border, apical tubular system, and supranuclear and infranuclear accumulations of glycogen. All other epithelial cell types were also preserved. The incorporation of [3H]thymidine and [3H]leucine continued during the culture period, reflecting a sustained synthesis of deoxyribonucleic acid and proteins. The hydrolytic activities of the brush border membrane were established based on data obtained throughout the course of the culture of a large number of intestinal specimens. Sucrase, maltase, glucoamylase, trehalase, lactase, alkaline phosphatase, and gamma-glutamyl transpeptidase activities increased during the 9 days of culture even though different patterns were recorded. These observations clearly established that human fetal small intestine can be maintained in organ culture for at least 9 days in a serum-free medium.
Asunto(s)
Intestino Delgado/embriología , Medios de Cultivo , Citoplasma/ultraestructura , ADN/biosíntesis , Retículo Endoplásmico/ultraestructura , Humanos , Intestino Delgado/metabolismo , Intestino Delgado/ultraestructura , Microscopía Electrónica , Mitosis , Técnicas de Cultivo de Órganos , Biosíntesis de ProteínasRESUMEN
Explants of suckling mouse jejunum have been maintained in serum-free organ culture with or without insulin added to the medium in order to determine the possible direct effect of this hormone on the hydrolytic functions of the brush border and on the proliferation of the crypt cells. The addition of insulin induced the precocious appearance of sucrase activity and increased trehalase, glucoamylase and lactase activities. Alkaline phosphatase activity remained unaffected in the tissue as well as in the medium. An increased DNA content and 3H-thymidine incorporation into DNA were already recorded after 24 h of culture. The mitotic index was significantly increased after 24 h and remained elevated when the culture was extended to 48 h. These results show that insulin directly influences the enzymatic maturation and the proliferation of intestinal epithelial cells of suckling mouse.
Asunto(s)
Insulina/farmacología , Mucosa Intestinal/efectos de los fármacos , Animales , Animales Lactantes , División Celular , ADN/biosíntesis , Enzimas/metabolismo , Mucosa Intestinal/crecimiento & desarrollo , Mucosa Intestinal/metabolismo , Yeyuno/efectos de los fármacos , Ratones , Ratones Endogámicos ICR , Técnicas de Cultivo de ÓrganosRESUMEN
The influence of hydrocortisone (10(-8)--10(-5) M) and thyroxine (10 (-9)--10(-6) M) on intestinal epithelial cell differentiation and proliferation have been studied using explants of suckling mouse jejunum maintained in serum-free organ culture. Hydrocortisone induced the appearance of sucrase activity and increased trehalase, glucoamylase, lactase and alkaline phosphatase activities. Thyroxine was completely ineffective at all the concentrations used. None of these hormones affected the mitotic activity or the 3H-thymidine incorporation into DNA. These results demonstrate that hydrocortisone but not thyroxine acts directly on intestinal brush border membrane differentiation and that both hormones do not influence the proliferation of the epithelial cells during postnatal development.