摘要:
目的 比较无血清条件下不同组织块法培养人角膜上皮细胞的出膜时间及出膜率.方法 培养方式:A组:剥除角膜内皮及刮除角膜上皮后组织块角膜上皮面向上贴壁;B组:剥除角膜内皮及刮除角膜上皮后组织块角膜上皮面向下贴壁;C组:仅剥除角膜内皮后全层组织块角膜上皮面向上贴壁;D组:仅剥除角膜内皮后全层组织块角膜上皮面向下贴壁.相差显微镜观察细胞形态学变化,对比各组出膜时间及出膜率.结果 A组:细胞爬出时间(7.00±3.00)d,出膜率47.50% (19/40);B组:细胞爬出时间(7.55±2.58)d,出膜率45.83% (11/24);C组:细胞爬出时间(8.43±3.32)d,出膜率63.64% (14/22);D组:细胞爬出时间(7.49±2.20)d,出膜率72.22% (39/54).卡方检验结果示,A组与D组、B组与D组出膜率相比,差异均有统计学意义(均为P <0.05),余组间两两比较差异均无统计学意义(均为P>0.05).结论 无血清条件下不同组织块培养方式细胞爬出时间大致相同,约一周.相比较而言,仅剥除角膜内皮后全层组织块角膜上皮面向下贴壁培养法出膜率明显较高,且培养的细胞更纯、更稳定.%Objective To compare and analyze the time of cell eruption formation and the rate of cell eruption of human corneal epithelial cells cultured by different explant culture methods under serum-free conditions.Methods Explant tissues were randomly divided into A,B,C and D groups.In A group,the cornea epithelial cells were put side up after stripping corneal endothelium and scraping the corneal epithelium;in B group,the corneal epithelium was put side down after stripping corneal endothelium and scraping the corneal epithelium;in C group,the corneal epithelium was put side up after only stripping corneal endothelium;and in D group,the corneal epithelium was put side down after only stripping corneal endothelium.The morphological changes of the cells in the 4 groups were observed by phase-contrast microscope,and both aforementioned variable were recorded.Results The average time of cell eruption formation and the rate of cell eruption in A,B,C and D group was (7.00 ± 3.00) d and 47.50% (19/40),(7.55 ±2.58)d and 45.83% (11/24),(8.43 ±3.32)d and 63.64% (14/22),(7.49 ± 2.20) d and 72.22% (39/54),respectively.The chi-square test showed that there was significant difference in the cell eruption rate between A and D group as well as B and D group (all P < 0.05),but there was no significant difference between the other groups (all P > O.05).Conclusion The time of cell eruption formation in the four different explant culture methods was about one week in free-serum conditions.And D group has higher cell eruption rate,and pure plus stable cultured cells when compared with the other three groups.