CAJ | 학술논문

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.方法研究经产妇知情同意,取P3代脐带间充质干细胞,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),用单因素方差统计分析增殖数量的差异.结果对照组细胞计数为(5.3±0.08)×106,每天开关箱5次,1、2和3min细胞计数分别为(4.5±0.14)×106、(3.9±0.08)×106和(3.27±0.19)×106,每天开箱10次,1、2和3min细胞计数分别为(3.48±0.06)× 106、(3.15±0.07)×106和(2.9±0.07)×106.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.
목적 탐토빈번개관배양상인기온도화CO2농도적변화대대규모배양제대간충질간세포(UC-MSCs)적영향.방법 연구경산부지정동의,취P3대제대간충질간세포,재37℃、5％ CO2배양조건하,모의조작중불동적개관배양상차수(매천0、5화10차),이급매차불동적개상시간(1、2화3min),용단인소방차통계분석증식수량적차이.결과 대조조세포계수위(5.3±0.08)×106,매천개관상5차,1、2화3min세포계수분별위(4.5±0.14)×106、(3.9±0.08)×106화(3.27±0.19)×106,매천개상10차,1、2화3min세포계수분별위(3.48±0.06)× 106、(3.15±0.07)×106화(2.9±0.07)×106.동양개상차수,수착매차개상시간적증장세포증식속도감만,1、2화3min각조세포증식수량차이균유통계학의의(F=265.0,P=0.000; F=1434.61,P=0.000)；동양개상시간조건하,매천개상10차조비5차조세포증식수감소,차이유통계학의의(t치위16.400,17.280,4.480,P균=0.000).수착개상시간적연장,개상차수증가,가이명현간도세포포장내출현공포화성편사망탈락적세포.결론 빈번개관배양상급개관상시간장단균가영향제대MSCs적증식,동시대세포형태유명현영향.
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 37C and 5％CO2.The door of the incubator was opened 0,5 or 10 times,each time for 1,2 or 3 min 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)×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)×106and (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.