RESUMO
<p><b>Objective</b>To explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HUMSCs) to differentiate into Leydig cells in the interstitial tissue of the rat testis.</p><p><b>METHODS</b>HUMSCs were obtained by tissue blocks culture attachment and their purity and multi-lineage differentiation ability were verified by flow cytometry and chondrogenic/adipogenic/osteogenic differentiation. Then the HUMSCs were marked by CM-Dil and transplanted into the interstitial tissue of the rat testis. At 4 and 8 weeks after transplantation, the survival and differentiation status of the HUMSCs were observed by immunofluorescence staining and flow cytometry. The suspension of the rat Leydig cells was obtained at 8 weeks for determining the expression of the Leydig cell marker 3β-HSD in the HUMSCs, the cells labeled with CM-Dil were sorted and cultured, and the medium collected after 3 days of culture for measurement of the testosterone level.</p><p><b>RESULTS</b>The expression of the Leydig cell marker CYPllal was not observed in the HUMSCs at 4 weeks but found at 8 weeks after transplantation and the differentiation rate of 3β-HSD was about 14.5% at 8 weeks. CM-Dil labeled cells survived after sorting and testosterone was detected in the medium.</p><p><b>CONCLUSIONS</b>HUMSCs are likely to differentiate into Leydig cells in the interstitium of the rat testis.</p>
Assuntos
Animais , Humanos , Masculino , Ratos , Biomarcadores , Metabolismo , Carbocianinas , Diferenciação Celular , Enzima de Clivagem da Cadeia Lateral do Colesterol , Metabolismo , Estudos de Viabilidade , Células Intersticiais do Testículo , Biologia Celular , Metabolismo , Células-Tronco Mesenquimais , Biologia Celular , Testículo , Biologia Celular , Fatores de Tempo , Cordão Umbilical , Biologia CelularRESUMO
<p><b>OBJECTIVE</b>To explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HuMSCs) to differentiate into Leydig cells through conditioned medium derived from Leydig cells.</p><p><b>METHODS</b>HuMSCs and Leydig cells were obtained by tissue blocks culture attachment and enzymatic digestion respectively. HuMSCs were induced by conditioned medium of Leydig cells as an experiment group while those before induction were cultured as a control group. The expressions of LHR, 3β-HSD and StAR in the induced HuMSCs were determined by RT-PCR after 3, 7 and 10 days of culture; those of CYP11A1, CYP17A1 and 3β-HSD measured by immunofluorescence staining after 2 weeks; and that of 3β-HSD detected by Western blot after 4 weeks.</p><p><b>RESULTS</b>The experimental group showed positively expressed LHR, 3β-HSD and StAR at 3, 7 and 10 days, CYP11A1, CYP17A1 and 3β-HSD at 2 weeks, and 3β-HSD at 4 weeks, while the control group revealed negative expressions at all the time points.</p><p><b>CONCLUSION</b>Induced with conditioned culture medium derived from Leydig cells, HuMSCs are likely to differentiate into steroidogenic cells and eventually into Leydig cells.</p>
Assuntos
Humanos , Masculino , Diferenciação Celular , Meios de Cultivo Condicionados , Células Intersticiais do Testículo , Biologia Celular , Células-Tronco Mesenquimais , Biologia Celular , Cordão Umbilical , Biologia CelularRESUMO
Leydig cells, located in the loose interstitial tissue of seminiferous tubules, are the major site for androgen synthesis and secretion, and play an important role in the reproductively and fertility of males. The dysfunction of Leydig cells may lead to various male diseases, such as primary hypogonadism, cryptorchidism, and hypospadias. This review outlines the recent findings concerning the generation, development and regulation of Leydig cells.