组织块重复贴壁使用在原代细胞培养中的实验意义

The Experimental Significance of Tissue Re-adherence Explant in Primary Cell Culture

目的旨在提高采用人脐静脉组织块贴壁法进行平滑肌细胞原代培养组织块的重复利用率,进而缩短采用细胞培养进行实验的培养周期。方法以改良人脐静脉平滑肌细胞贴壁培养法为基础,当原代培养细胞(对照组)进行第1次细胞传代后,将培养组织块再次接种于新培养瓶中继续培养(实验组),分别记录两组细胞爬出时间、第1次传代时间、以及第1次传代后24、48、72、96h的细胞计数,通过倒置显微镜对两组细胞进行形态学观察,采用抗α-actin免疫细胞化学方法,对两组传代细胞的类型和性质进行鉴定。结果与对照组比较,实验组细胞爬出时间及第1次传代时间均明显缩短(P<0.05);两组第1次传代后24、48、72、96h细胞计数差异均无统计学意义(P>0.05);两组细胞形态经α-actin鉴定差异无统计学意义。结论人脐静脉组织块重复贴壁培养后,经传代其细胞数量、细胞形态与原代组织块贴壁培养的细胞差别均无统计学意义。

Objective To improve the reuse of smooth muscle tissue during primary culture in human umbili- cal vein tissue re - explant method, and to shorten the cycle of cell culture. Methods Based on modified hu- man umbilical vein tissue explant method,when primary culture cells fused 80% and proceeded the first gener- ation cell passage（ Control group）, the cultured tissue were explanted and cultured in another new flask （ex- perimental group）. The cells of two groups were observed using inverted microscopes for morphological fea- tures. Climbing time of smooth muscle cells in two groups and the counting of cell after the first passage 24, 48,72,96h were recorded. Human umbilical vein smooth muscle cells were identified by a-actin. Results Compared with the control group, the time of climbing and the first passage were reduced significantly in exper- imental group（ P 〈0.05 ）. There was no significant difference in counting cells after the first passage 24,48, 72,96h between two groups（P 〉 0.05 ）. Conclusion It is no difference in number of ceils, morphology be- tween re - explanting culture and primary cell culture after re - explanting culture of human umbilical vein tis- sue.