Lin Fan-Li, Qian Yi, Yang Min, Wang Shu-Yue, Chen Xiao-Qing, Cheng Yan, Xiong Hao, Huang Chun-Lan
Department of Hematology, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan Province, China.
Department of Hematology, The Affiliated Hospital of Southwest Medical University, Luzhou, 646000, Sichuan Province, China,E-mail:
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2019 Apr;27(2):594-599. doi: 10.19746/j.cnki.issn1009-2137.2019.02.046.
To investigate the effect of microvascular endothelial cells (MEC) on the proliferation of hematopoictic stem cells (HSC) under different culture conditions in vitro.
The MEC from lung tissue of C57BL/6 mice and the HSC from bone marrow of GFP mice were used for non-contact co-culture, 2 D contact co-culture, at same time the single MEC and single HSC culture were seted up and were used as control group. The cell counting and CCK-8 method were used to detect and compare the proliferation levels of suspension cells in different groups on day 1, 3, 5 and 7.
MEC presented adherent growth. In process of cell culture in vitro, the number of suspension cells in MEC and HSC co-culture group and single HSC culture group increased, the suspension cells in 2D contact and non-contact co-culture groups more early gated into logarithmic growth phase as compared with suspension cells in control group, the proliferation level of suspention cells in 2D contact culture group was higher than that in non-contact co-culture group and single HSC culture group (P<0.05), the proliferation level of suspension cells in non-contact co-culture group was higher than that in single HSC culture group (P<0.05).
The culture of HSC in vitro can proliferate HSC, MEC can promote the proliferation of HSC, MEC also can promote the HSC proliferation by non-contact co-culture in vitro, which relates with the effect of cytokines secreted from MEC; the effect of MEC and HSC contact co-culture on the proliferation of HSC is stronger than that of non-contact co-culture, which relates with the regulation of cell-cell contact.
研究体外不同培养条件下微血管内皮细胞（MEC）对造血干细胞（HSC）增殖的影响。
采用C57BL/6小鼠肺组织的MEC和绿色荧光蛋白（GFP）小鼠骨髓的HSC进行非接触共培养、二维接触共培养，同时设置MEC单培养组和HSC单培养组作为对照组。采用细胞计数和CCK-8法检测并比较第1、3、5和7天不同组悬浮细胞的增殖水平。
MEC呈贴壁生长。在体外细胞培养过程中，MEC与HSC共培养组和HSC单培养组悬浮细胞数量增加，二维接触共培养组和非接触共培养组悬浮细胞较对照组更早进入对数生长期，二维接触共培养组悬浮细胞增殖水平高于非接触共培养组和HSC单培养组（P<0.05），非接触共培养组悬浮细胞增殖水平高于HSC单培养组（P<0.05）。
体外培养HSC可使其增殖，MEC可促进HSC增殖，MEC还可通过体外非接触共培养促进HSC增殖，这与MEC分泌的细胞因子作用有关；MEC与HSC接触共培养对HSC增殖的促进作用强于非接触共培养，这与细胞间接触的调节有关。
