CAJ | 학술논문

目的观察全层人胚胎视网膜的短期、中期及长期保存后的活性和结构.方法 12～24周全层人胚胎视网膜22例分为88片,明胶包被后分别行Ames液、中期保存液(DX角膜保存液)、-80℃、程序深低温4种方法保存不同时间后,锥虫蓝染色观察视网膜神经上皮层活细胞百分率,光学显微镜和透射电子显微镜观察视网膜组织结构.结果人胚胎视网膜在新鲜取材时活细胞占(94.79±2.85)%;短期保存在Ames液4 h内活细胞百分率大于80%,中期保存在DX液1～2 d时活细胞百分率大于77%,均与新鲜取材时活细胞百分率无显著差异.-80℃保存7 d时活细胞百分率为(65.83±5.06)%,1个月时降至(57.54±16.18)%;深低温长期保存1个月时活细胞百分率为(69.46±9.31)%.全层人胚胎视网膜保存在Ames液2 h时光学显微镜和电子显微镜检查均未见明显异常;在DX液中2 d、-80℃7 d和深低温1个月时光学显微镜检查见其层次清楚,但细胞间隙加大,组织排列疏松.结论全层人胚胎视网膜在Ames液和DX角膜保存液中可于一定时间内保持较好的活性和组织结构.
목적 관찰전층인배태시망막적단기、중기급장기보존후적활성화결구.방법 12～24주전층인배태시망막22례분위88편,명효포피후분별행Ames액、중기보존액(DX각막보존액)、-80℃、정서심저온4충방법보존불동시간후,추충람염색관찰시망막신경상피층활세포백분솔,광학현미경화투사전자현미경관찰시망막조직결구.결과 인배태시망막재신선취재시활세포점(94.79±2.85)%;단기보존재Ames액4 h내활세포백분솔대우80%,중기보존재DX액1～2 d시활세포백분솔대우77%,균여신선취재시활세포백분솔무현저차이.-80℃보존7 d시활세포백분솔위(65.83±5.06)%,1개월시강지(57.54±16.18)%;심저온장기보존1개월시활세포백분솔위(69.46±9.31)%.전층인배태시망막보존재Ames액2 h시광학현미경화전자현미경검사균미견명현이상;재DX액중2 d、-80℃7 d화심저온1개월시광학현미경검사견기층차청초,단세포간극가대,조직배렬소송.결론 전층인배태시망막재Ames액화DX각막보존액중가우일정시간내보지교호적활성화조직결구.
Objective To observe the configuration and viability of full thickness human fetal retinaafter short-,mid-and long-term preservation.Methods Twenty-two full thickness human fetal retinaeof gestational age of 12-24 weeks were coated by glutin and cut into 88 pieces,and then preserved inAmes' solution,DX solution,-80℃ refrigerator or under cryopreservation condition.The cell viability ofretinal neuroepithelial layer was determined by trypan blue staining,retinal configuration was determinedby light microscope and electromicroscope.Results The viability of neuroepithelial layer was(94.79±2.85)% in fresh fetal retina,>80% in Ames' solution within 4 hours,and>77% in DX solution within2 days.There was no significant difference between those solution-preservations and the fresh fetal.In-80℃ refrigerator,the viability was(65.83±5.06)% after 7 days,and then dropped tO(57.54±16.18)% at the end of the first month.Under the cryopreservation condition,the viability was(69.46±9.31)% at the end of first month.Light and transmission electron microscopy had not deteced anyabnormals in the full thickness human fetal retina preserved in Ames's solution within 2 hours,but showedclear 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' solutionand DX solution can preserve good viability and configuration of full thickness human fetal retina in acertain time period.