角膜缘基质诱导毛囊干细胞角膜上皮样转分化的研究  

Transdifferentiation of rat hair follicle stem cells to epithelium-like cells induced by stroma of limbus corneae

作　　者：姜自玲[1] 杨力[1] 连小华[2] 杨恬[2] 杨柯[2] 

机构地区：[1]重庆大学生物工程学院国家“111计划”基地,重庆400044 [2]第三军医大学基础医学部细胞生物学教研室,重庆400038

出　　处：《第三军医大学学报》2008年第13期1235-1238,共4页Journal of Third Military Medical University

基　　金：高等学校学科创新引智计划(“111”)(B06023);国家自然科学基金(30671888);重庆市科委自然科学基金(2003BB5007)~~

摘　　要：目的体外诱导培养毛囊干细胞角膜上皮样分化。方法体外分离培养毛囊干细胞,然后用角膜缘基质组织匀浆液对其进行诱导培养,并用免疫细胞化学(ICC)法检测诱导前后α6-integrin、CD34、K12的变化;RT-PCR方法检测细胞诱导前后K12的mRNA表达情况;流式细胞分析(FCM)检测毛囊干细胞角膜上皮样分化的阳性率。结果体外培养的毛囊干细胞保持高增殖的状态,α6-integrin、CD34于细胞质表达。经过10d的诱导培养,可见K12于细胞质阳性表达;RT-PCR结果显示毛囊干细胞经诱导后K12的mRNA明显表达;FCM结果亦显示毛囊干细胞经诱导培养后有较高比率的K12阳性细胞。结论毛囊干细胞经角膜缘组织匀浆液体外诱导可成功地分化为角膜上皮样细胞。Objective To explore how to induce the hair follicle stem cells differentiate into epitheliumlike cells, Methods The hair follicle stem cells of 7- to 8-day SD rats were isolated and cultured, then induced by homogenate of rabbit corneal limbal stroma. The expressions of α 6-integrin, CD34 and K12 were detected by immunocytochemistry （ICC）. The mRNA change of Keratin-12（K12） was also detected by RT-PCR. The positive rate of differentiation was detected by flow cytometry （FCM）. Results The cultured hair follicle stem cells were always in high proliferation, α 6-integrin and CD34 were expressed in the cytoplasm, K12 expressed in the cytoplasm after being induced for 10 d. The mRNA of K12 was also expressed obviously. And a considerable rate of the K12-positive cells was also observed by FCM. Conclusion Induced by homogenate of limbus corneae, hair follicle cells can differentiate into corneal epithelium-like cells in vitro.

关 键 词：毛囊干细胞 角膜上皮细胞 转分化 

分 类 号：R322.91[医药卫生—人体解剖和组织胚胎学] R329-33[医药卫生—基础医学]