温度和二氧化碳变化对大规模培养脐带间充质干细胞的影响  

Effect of temperature and concentration of carbon dioxide on large-scale culturing of human umbilical cord mesenchymal stem cells

作　　者：林娜[1] 陈津[1] 付云烽[1] 施小华[1] 王庆华[1] 谭建明[1] 

机构地区：[1]南京军区福州总医院全军器官移植中心,福州350025

出　　处：《中华细胞与干细胞杂志（电子版）》2012年第1期14-16,共3页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：国家自然科学基金(30571759);全军"十五"重大课题(04Z007);福建省科技厅重大攻关课题(2002Y007)

摘　　要：目的 探讨频繁开关培养箱引起温度和CO2浓度的变化对大规模培养脐带间充质干细胞(UC-MSCs)的影响。方法 取P3代UC-MSCs,在37℃、5﹪?CO2培养条件下,模拟操作中不同的开关培养箱次数(每天0、5和10次),以及每次不同的开箱时间(1、2和3min),对比分析细胞生长形态和增殖数量的差异。结果 同样开箱次数,随着每次开箱时间的增长细胞增殖速度减慢,1、2和3min各组细胞增殖数量差异均有统计学意义(F=265.0,P=0.000;F=1434.61,P=0.000);同样开箱时间条件下,每天开箱10次组比5次组细胞增殖数减少,差异有统计学意义(t值为16.400,17.280,4.480,P均?=0.000)。随着开箱时间的延长,开箱次数增加,可以明显看到细胞胞浆内出现空泡和成片死亡脱落的细胞。结论 频繁开关培养箱及开关箱时间长短均可影响脐带MSCs的增殖,同时对细胞形态有明显影响。Objective To explore the effect of temperature and carbon dioxide（CO2） concentration fluctuation caused by opening the incubator on the growth of human umbilical cord derived mesenchymal stem cells （UC-MSCs）. Methods The UC-MSCs （3rd passage） were cultured at 37℃ and 5 % CO2. The door of the incubator was opened 0, 5 or 10 times,each time for 1, 2 or 3 rain respectively every day. Results The cell count of normal control is （5.3 ± 0.08）×106. When the incubator was opened for 5 times, 1, 2, and 3 min for each time, the cell numbers were （4.5 ± 0.14） x× 106, （3.9 ±0.08） × 106 and （3.27 ± 0.19） ×106 respectively. When the incubator was opened for 10 times, 1, 2, and 3 min for each time, the cell numbers were （3.48 ±0.06） × 106. （3.15 ± 0.07） ×106 and （2.9± 0.07）× 106 respectively. For a given number of door opening, there was statistical significance for cell numbers when the duration of opening was 1, 2 or 3 min （F= 265.0, P = 0.000; F= 1434.61, P = 0.000） ,The growth of UC-MSCs significantly slowed down as the frequency of door opening increased. When the incubator was opened for 10 times, cell growth significantly decreased compared with that for 5 times （t -- 16.400, t = 17.280, t = 4.480, P =0.000）. In addition the morphology and viability of cells were worsened when the incubator was opened for a longer duration.Conclusion The study demonstrated that frequent opening of the incubator may slow down the proliferation of UC-MSCs and have a deteriorating effect on cell morphology and viability.

关 键 词：脐带间充质干细胞 温度 二氧化碳 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]