Objective To evaluate the feasibility of preservation of arteriesby vitrification and the effectiveness of vitrified arteries as allografts. Methods Sixty rabbits were used in the research. Forty-eight femoral arteries wereharvested from 24 rabbits as the transplanted materials，and 24 femoral arteries were preserved by vitrification, 24 by freezing for 14 days，respectively. Theother 36 rabbits were used as the transplanted subjects，and were divided into three groups， 12 rabbits including 24 femoral arteries per group: Group A(fresharterial autografts), Group B(vitrified arterial allografts) and Group C(frozen arterial allografts). The morphologic changes of arterial grafts were observed macroscopically and histologically. The patent rate of arterial grafts were measured by angiography, and the rabbits were sacrificed on the 14th day, the 30th day, the 60th day and the 120th day after transplantation respectively. Arterial grafts were harvested to observe the morphological changes，and the immunological rejection was evaluated by measuring the ratio of tunica intima and tunica media. The results were compared between these groups. Results Before transplantation，theintegrated rate of Group B was 91.67%，which was significantly better than that of Group C（54.17%, Plt;0.01）. After transplantation, the accumulative patent rate of Group B was 87.50%，which was significantly better than that of Group C（66.67%, Plt;0.05）. There was statistically significant difference in the ratioof tunica intima and tunica media between Group B and Groups A, C（Plt;0.05）.Conclusion The above results show that vitrification does less damage to cells and tissues because of ice-free in the process of cryopreservation. So vitrification can be used to preserve arteries， and the arterial allografts preserved by vitrification are better than those preserved by freezing.
Objective To observe the configuration and viability of full thickness human fetal retina after short-, mid- and long-term preservation. Methods Twenty-two full thickness human fetal retinae of gestational age of 12-24 weeks were coated by glutin and cut into 88 pieces, and then preserved in Ames' solution, DX solution, -80℃ refrigerator or under cryopreservation condition. The cell viability of retinal neuroepithelial layer was determined by trypan blue staining, retinal configuration was determined by light microscope and electromicroscope. Results The viability of neuroepithelial layer was (94.79plusmn;2.85) % in fresh fetal retina, gt;80% in Ames' solution within 4 hours, and gt;77% in DX solution within 2 days. There was no significant difference between those solution-preservations and the fresh fetal. In -80℃ refrigerator, the viability was (65.83plusmn;5.06)% after 7 days, and then dropped to (57.54plusmn;16.18)% at the end of the first month. Under the cryopreservation condition, the viability was (69.46plusmn;9.31)% at the end of first month. Light and transmission electron microscopy had not deteced any abnormals in the full thickness human fetal retina preserved in Ames' solution within 2 hours, but showed clear retinal layers with bigger intercellular space after preserved in DX solution for 2 days, in -80℃ refrigerator for 7 days and under cryopreservation condition for 1 month. Conclusion Ames' solution and DX solution can preserve good viability and configuration of full thickness human fetal retina in a certain time period.
PURPOSE:To establish methods for cryopreservation of human retinal pigment epithelial cells (RPEs)and cell culture from thawing of frozen cells. METHODS:Primary cultured RPEs or its first or second passages,added with 10 dimetbylsulfoxide,were kept in --20℃ for 1 to 2 hours,and then further froze to -40~C over night before being placed in liquid nitrogen. The frozen cells were thawed in 60℃ within 2 minutes. Trypan blue staining and immunocytochemical staining with anti-human keratin were performed for cell viability and differentiation. The growth curve was also determined by calculating the total number of cells/well/day. RESULTS:The viable rate from frozen RPEs was 90%. No differences were observed for growth activity between cultures from frozen cells and controls. The cells were positive with anti-human keratin staining. The logarithmic growth phase was during I to 4 days and the doubling time yeas 1.55 days. CONCLUSION: Cryopreservation of RPEs in liquid nitrogen can maintain biological activities of cells with normal growth and features after thaw- ing. This will provide cell lines for in vitro experiments and possibly for cell banks for RPE transplantation for some fundus diseases. （Chin J Ocul Fundus Dis,1997,13:157-159）