欢迎访问《解剖学报》官方网站！今天是
English
小鼠睾丸支持细胞对人脐带间充质干细胞增殖能力的影响
Effect of mouse Sertoli cells on human umbilical cord mesenchymal stem cells proliferation
目的 探讨睾丸支持细胞（SCs）对体外培养人脐带间充质干细胞(hUCMSCs）增殖能力的影响。方法 体外分离培养并鉴定hUCMSCs和SCs；
Transwell-insert系统共培养SCs 和hUCMSCs（共培养组）；普通DMEM-F12培养基培养的hUCMSCs作为对照（对照组）；添加细胞因子
(白细胞介素-3，粒细胞-巨噬细胞集落刺激因子) 的DMEM-F-12培养基培养的hUCMSCs作为阳性对照 (细胞因子组）。用流式细胞术、免疫细胞
化学等方法检测SCs对hUCMSCs增殖、细胞周期、表面标记分子表达的影响，透射电镜观察细胞超微结构。结果 与对照组相比，共培养组和细胞
因子组hUCMSCs增殖明显加快，以共培养组尤为显著（P<0.05）；共培养组和细胞因子组hUCMSCs 表面分子CD29和CD105阳性率均增高；细胞周
期分析显示，共培养组G0/G1期细胞数与对照组相比无明显改变，而细胞因子组略有减少。透射电镜观察对照组与共培养组细胞均呈现原始细胞
的超微结构特点。结论 SCs共培养可促进hUCMSCs增殖，且共培养后仍保持其干细胞特性。
Objective To observe the regulatory effect of Sertoli cells (SCs) on proliferation in human umbilical cord mesenchymal
stem cells (hUCMSCs) in vitro. Methods hUCMSCs and SCs were isolated, cultured and identified in vitro. A co-culture system was
established by culturing SCs in the Transwell insert and hUCMSCs on the plastic plates(the co-culture group). hUCMSCs culture in
DMEM-F12 medium was used as the control group. hUCMSCs culture in DMEM-F12 medium supplemented IL-3 and GM-CSF served as the
cytokine group. Proliferation, cell cycle analyse and surface marker molecules of hUCMSCs were studied by flow cytometry and
immunocytochemistry. Ultrastructures of the cultured cells were observed by transmission electron microscopy. Results Compared
with the control, proliferation of hUCMSCs in the co-culture- and cytokine-group increased significantly in the co-culture group
(P<0.05). The expression of CD29 and CD105 on the surface of hUCMSCs in the co-culture and the cytokine group also increased.
Cell cycle analysis showed that there were a few cells arrested at quiescent phases in the co-culture and cytokine groups,
especially in the cytokine group. In the coculture group and the control group，hUCMSCs may displayed the ultrastructural features
of the primitive cells by transmission electron microscopy. Conclusion Proliferation of hUCMSCs may be promoted as co-culture with
SCs and still kept their stem cell properties after the co-culture.
脐带间充质干细胞 / 支持细胞 / 共培养 / 体外培养 / 免疫组织化学 / 流式细胞术 / 小鼠
Umbilical cord mesenchymal stem cell / Sertoli cell / Co-culture / Culture in vitro / Immunohistochemistry / Flow cytometry / Mouse
［1］Wang HS, Hung SC, Peng ST, et al. Mesenchymal stem cells in the Wharton’s jelly of human umbilical cord ［J］. Stem Cells, 2004, 22(7): 1330-1337.
［2］Chao KC, Yang HT, Chen MW. Human umbilical cord mesenchymal stem cells suppress breast cancer tumorigenesis through direct cell-cell contact and internalization ［J］. J Cell Mol Med, 2012, 16(8):1803-1815.
［3］Kim JY, Jeon HB, Yang YS, et al. Application of human umbilical cord blood-derived mesenchymal stem cells in disease models ［J］. World J Stem Cells, 2010, 2(2): 34-38.
［4］Othberg AL, Willing AE, Cameron DF, et al. Trophic effect of porcine Sertoli cells on rat and human ventral mesencephalic cells and hNT neurons in vitro ［J］. Cell Transplant, 1998, 7(2): 157-164.
［5］Halberstadt C, Emerich DF, Gores P. Use of Sertoli cel1 transplants to provide local immunoprotection for tissue grafts ［J］. Expert Opin Biol Ther, 2004, 4(6):813-825.
［6］Emerich DF, Sanberg PR.Novel means to selectively identify sertoli cel1 transplants ［J］.Cell Transplant, 2002, 11(6): 507-512
［7］Zucconi E, Viera NM, Bueno DF, et al. Mesenchymal stem cells derived from canine umbilical cord vein-a novel source for cell therapy studies［J］. Stem Cell Dev, 2010, 19(3): 395-402.
［8］Wei X, Peng G, Zheng S, et al. Differentiation of umbilical cord mesenchymal stem cells into steoidogenic cells in comparison to bone marrow mesenchymal stem cells ［J］. Cell Prolif, 2012, 45(2): 101-110.
［9］Zhang DY, He DW, Wei GH, et al. Long-term coculture of spermatogonial stem cells on Sertoli cells feeder layer in vitro［J］. Sichuan Da Xue Xue Bao Yi Xue Ban, 2008, 39(1):6-9.
［10］Luca G, Calafiore R, Basta G, et al. Improved function of rat islets upon co-microencaspsulation with Sertoli’s in alginate/poly-L-ornithine［J］. AAPS PharmSci Tech, 2001, 2(3): E15.
［11］Fan XB, Liu TQ, Hao YJ, et al. Optimization for dissociation and culture of mesenchymal stem dells derived drom umblical cord blood ［J］. Progress in Biochemistry and Biophysics, 2008, 35(8):905-913. (in Chinese)
范秀波，刘天庆，郝永杰，等. 脐带血来源间充质干细胞体外分离培养条件的优化 ［J］. 生物化学与生物物理进展, 2008, 35(8):905-913.
［12］Fan X, Liu T, Liu Y, et al. Opitimization of primary culture condition for mesenchymal stem cells derived from umbilical cord blood with factorial sedign ［J］. Biotechnil Prog, 2009, 25(2): 499-507.
［13］Dyer KD, Moser JM, Czapiga M, et al. Functionally competent eosinophils differentiated ex vivo in high purity from normal mouse bone marrow ［J］. J Immunol, 2008, 181(6): 4004-4009.
［14］Leon CM, Barbosa CM, Justo GZ, et al. Requiement for PLCy2 in IL-3 and GM-CSF-stimulated MEK/ERK phosphorylation in murine and human hematopoietic stem/progenitor cells［J］. J Cell Physiol, 2011, 226(7): 1780-1782.
［15］Fan P, He L, Pu D, et al.Testicular Sertoli cells influence the proliferation and immunogenity of co-cultured endothelial cells［J］. Biochem Biophys Res Commun, 2011, 404 (3): 829-833.
［16］Lu YM, Fu WY, Piao YJ, et al. The ultrastructure of human mesenchymal stem cells［J］. Journal of Chinese Electron Microscopy Society, 2002,21(4):373-376. (in Chinese)
路艳蒙, 傅文玉, 朴英杰, 等.人骨髓间充质干细胞的超微结构［J］.电子显微学报2002,21(4):373-376.
［17］Zhao J, Jin J, Liang YJ, et al. Effect of Sertoli cells on mouse embryonic cerebral cortical cells in vitro［J］. Journal of Clinical Rehabilitative Tissue Engineering Research, 2008, 12(43):8490-8494. (in Chinese)
赵佳，金洁，梁元晶，等. 睾丸支持细胞对体外培养鼠胎脑皮质细胞的影响［J］. 中国组织工程研究与临床康复, 2008, 12, (43):8490-8494.
贵州省社会发展科技攻关项目 黔科合SY[2009]3071；贵州省教育厅项目，黔教高发[2010]305;省自然科学基金资助项目
/
| 〈 |
|
〉 |
