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[A study on effect of basic fibroblast growth factor on human limbal stem cell proliferation cultured in low calcium
1Department of Ophthalmology, the First Hospital, Peking University, Beijing 100034, China.
Objective:
To study how to obtain relatively pure stem cells and the effects of bFGF on the human limbal stem cell proliferation in order to set up the foundation of cultured limbal stem cells for treatment of ocular surface and tear diseases.
Methods:
Limbal button inoculation was used in low calcium culture. Monoclonal antibody AE5 was used to identify the stem cells. To examine the effects of bFGF (1 - 100 ng/ml) on stem cell proliferation, the technique of digital camera and computer image analysis system were used.
Results:
Cultured cells were mostly corneal stem cells. This was the first time to combine the method of obtaining basal cell layer and culture with low calcium medium, which could harvest relatively pure and undifferentiated stem cells in localization and nature. The basic FGF (1 - 100 ng/ml) promoted cultured stem cells to proliferate significantly (P < 0.001).
Conclusions:
Limbal stem culture provides resources of stem cells. It is very useful in treatment of ocular surface and tear diseases. The basic FGF plays an important role in proliferation of stem cells in graft. Computer image analysis system is an effective and simple way for evaluating the proliferation of a primary culture.