| Purpose: To isolate and culture corneal endothelial cells (CECs) and Mesenchymal stem cells (MSCs) deprived from marrow of New Zealand white rabbits and observe their growth status in vitro .Then evaluate the feasibility of CECs transplantation by a Gelatin membrane carrier and replaced by autologous MSCs. Also,to observe the short-term growth status and function of CECs after transplantation and to observe the differentiation and function of MSCs after they were transplanted into the CECs position in vivo.Method: For CECs culture, the Descemet' s membrane and endothelial cell layer of rabbits cornea were torn apart and cut into small pieces , then trypsined by trypsin-EDTA solution. The isolated CECs were cultured subsequently. For MSCs culture , the bone marrow was aspirated from iliac of the rabbits. Then the MSCs were isolated by density centrifugation and cultured. The cells were observed under the microscope everyday. And the MSCs were identified by immunocytochemical method with mouse antibody to rabbits CD44 and CD45, which were labelled separately by FITC and PE, then checked by flow cytometer. The two types of cells were also labelled by Brdu and identified by anti-Brdu antibody. CECs of the first passage were grown in vitro on glutaraldehyde-fixed Gelatin membranes and then the membranes were glued by a -cyanoacrylate alkyl to 7.00mm autologous rabbits' corneal bottons whose endothelium were mechanically removed previously. Then the botton were sutured in place. By the same way autologous MSCs were transplanted into the CECs place to replace the CECs . And the controls were transplanted Gelatin membranes without any cells. The rabbits were breeded and observed by slit microscope and confocal microscopy at different intervals postoperation. Also , IOP and corneal thickness were measured by Perkin' s tonometer and ultrasonic pachymeter . Results: CECs of high purity were isolated by this method, The cells showed contact...
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