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目的:探索角膜基质接触诱导胚胎干细胞(embryonic stem cells,ES.细胞)定向分化的可能性。方法:分别在去上皮的新西兰白兔表层角膜缘基质上、晶状体上皮细胞饲养层上培养ES-D_3细胞,裸鼠皮下移植使其形成复层细胞,一段时间后,行扫描电镜和光镜观察。结果:I.ES细胞在新鲜表层角膜缘基质上增殖缓慢,2～3周形成较大细胞集落,光镜下细胞形态单一,体积较正常ES细胞大,电镜下细胞核已演变为细长形,移植到裸鼠皮下2周形成上皮样细胞复层,电镜下可见微绒毛,而角膜基质非上皮面的ES细胞仍保持其小细胞状态不变。2.晶状体上皮细胞与ES细胞共培养只能延缓其分化时间,不能诱导其定向分化。结论:表层角膜缘基质具有诱导ES细胞定向分化的潜能,体外培养条件下可能不发生晶状体诱导的第三级胚胎诱导。眼科学报1999;15:195-198。
Objective: To explore the possibility of corneal stromal contact induced differentiation of embryonic stem cells (ES cells). Methods: ES-D 3 cells were cultured on the corneal limbal epithelium of New Zealand white rabbits, respectively. The nude mice were transplanted subcutaneously to form the stratum corneum. After a period of time, ES-D 3 cells were observed by scanning electron microscopy and light microscopy. Results: The proliferation of I. ES cells was slow on fresh corneal limbal stroma, forming large cell colonies after 2 ~ 3 weeks. Under the light microscope, the cell morphology was single and its volume was larger than that of normal ES cells. Under the electron microscope, the nucleus had evolved into slender, Transplanted into nude mice subcutaneously for 2 weeks to form epithelial cell layer, microvilli can be seen under electron microscope, while the non-epithelial ES cells in corneal stroma keep their small cell state unchanged. Coculture of lens epithelial cells with ES cells can only delay their differentiation time, and can not induce their directional differentiation. CONCLUSIONS: The limbal stromal matrix has the potential of inducing the directional differentiation of ES cells and may not induce lens-induced tertiary embryogenesis in vitro. Journal of Ophthalmology 1999; 15: 195-198.