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[Preliminary experimental study on commitment differentiation of embryonic stem cells induced by corneal limbal stroma in vitro].

PURPOSE
To investigate the possibility of commitment differentiation of embryonic stem cells (ES cells) induced by corneal limbal stroma.


METHODS
Superficial corneal limbal stroma and lentis epithelium from New Zealand white rabbit were used for feeder layer for ES-D3 cells culture to induce the ES cells differentiation into corneal limbal stem cells. The differentiated ES cells were determined by light microscope and electronic microscope.


RESULTS
1. The proliferation of differentiating ES cells which cultured on corneal surface stroma layer around the corneal limbus was slow. The appearance of induced ES cells were uniform cells with much larger size than undifferentiated ES cells, and the nucleoli become long and thin under electronic microscope, but the size of ES cells cultured on deep stroma maintain as small as undifferentiated ES cells. The ES cells cultivated on surface stroma layer around the corneal limbus appeared microvilli 2 weeks after grafting into subcutaneous of athymic mice. 2. Lentis epithelium can only delay but not induce the ES cells differentiation to corned limbal stem celluloid.


CONCLUSION
These results suggest that the superficial corneal limbal stroma has a potential ability to induce ES cells commitment differentiation. The tertiary embryo induction may not occur in vitro.