摘要
目的探讨内毒素(Endotoxin),又称为脂多糖(lipopolysaccharide,LPS)的量效和时效对人脐带间充质干细胞(human umbilical cord mesenchymal stem cells,hUC-MSCs)增殖的影响。方法用含不同浓度LPS(0.098、0.195、0.39、0.78、1.56、3.12、6.25、12.5、25和50 EU/mL)的干细胞培养基培养hUC-MSCs,采用CCK-8增殖实验检测24h、48h和72 h的细胞增殖情况。确定促进hUC-MSCs增殖的低浓度(0.098 EU/mL)、中浓度(1.56 EU/mL)和高浓度(25 EU/mL)LPS,选择作用时间72 h,进一步用流式细胞术明确LPS对hUC-MSCs增殖的影响。结果CCK-8检测结果提示,作用24 h、48 h时,表现为抑制hUC-MSCs的增殖。作用72 h时,表现为促进细胞增殖,且低浓度区域促进作用最显著,浓度增加其促增殖作用相对减弱。流式细胞术结果提示低浓度LPS组G0/G1期的占比降低,S期和G2/M期占比增高。结论LPS的量效和时效对hUC-MSCs的增殖表现出不同的效应,适宜的LPS浓度及作用时间有利于hUC-MSCs的增殖。
Objective To investigate the effects of dose-response and time-dependent of endotoxin on the proliferation of human umbilical cord mesenchymal stem cells(hUC-MSCs).Methods The hUC-MSCs were cultured in stem cell culture medium containing different concentrations of endotoxin(0.098,0.195,0.390,0.780,1.5600,3.12,6.250,12.500,25.000 and 50.000 EU/mL).CCK-8 proliferation assay was used to detect the cell proliferation at 24,48 and 72 hours.The low concentration(0.098 EU/mL),medium concentration(1.56 EU/mL)and high concentration(25 EU/mL)of LPS were determined to promote the proliferation of hUC-MSCs,and the action time was selected for 72 hours.The effect of LPS on the proliferation of hUC-MSCs was further determined by flow cytometry.Results After 24,48 hours of the treatment,it was mainly manifested by inhibiting the proliferation of h UC-MSCs.After 72 hours of the treatment,it showed a trend of promoting cell proliferation,and the promotion effect was most significant in the low concentration area,and the promotion effect was relatively weakened when the concentration increased.Further flow cytometry results suggested that the proportion of G0/G1phase decreased,S phase and G2/M phase increased in LPS group of low concentration.Conclusion The doseresponse and time-dependent of endotoxin have different effects on the proliferation of h UC-MSCs,and the appropriate concentration and duration of action of endotoxin are beneficial to the proliferation of h UC-MSCs.
作者
袁伟
孟明耀
李欣欣
熊晶晶
李檬
曹佳
刘梅
侯宗柳
黄永坤
YUAN Wei;MENG Mingyao;LI Xinxin;XIONG Jingjing;LI Meng;CAO Jia;LIU Mei;HOU Zongliu;HUANG Yongkun(Dept,of Pediatrics,The 1st Affiliated Hospital ofKunming Medical University,Kunming Yunnan 650032;Yunnan Provincial Key Laboratory of Tumor Immunotherapy,Kunming Yunnan 650051;Yan'an Hospital Affiliated to Kunming Medical University,Kunming Yunnan 650051;Yunnan Provincial Key Laboratory of Laboratory Medicine,Kunming Yunnan 650032,China)
出处
《昆明医科大学学报》
CAS
2022年第9期7-12,共6页
Journal of Kunming Medical University
基金
云南省检验医学重点实验室开放课题基金资助项目(2017DG005)。
关键词
内毒素
人脐带间充质干细胞
细胞增殖
干细胞移植
Endotoxin
Human umbilical cord mesenchymal stem cells
Cell proliferation
Stem cell transplantation
