Na Xiao-dong, Zhao Zi-ping, Tan Meng-qun, Xie Qi-yang, Wang Qi-ru
Research Laboratory of Blood Physiology, Xiang Ya Medical College of Central South University, Changsha 410078, China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2002 Feb;24(1):36-40.
To investigate the effects of murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of yolk sac hematopoietic progenitors.
The serum-free mBMEC-CM was obtained from subcultures of murine endothelial cell line derived from bone marrow which was established in our laboratory. The murine yolk sacs were harvested on day 8.5 postcoitus (pc) and incubated with 0.1% collagenase in 10% fetal calf serum at 37 degrees C for 40 minutes. Yolk sac cells were incubated in tissue culture dishes at 37 degrees C for 1 hour. Nonadherent cells were collected for semisolid culture assay of granulocyte-macrophage colony forming unit (CFU-GM) and high proliferative potential-colony forming cell (HPP-CFC) after being cultured in DMEM with 10% mBMEC-CM and 10% FBS for 24 hours. The number of CFU-GM and HPP-CFC was counted at day 7 and day 14 respectively.
The growth of CFU-GM and HPP-CFC was supported by mBMEC-CM with GM-CSF. mBMEC-CM could induce the proliferation and differentiation of yolk sac hematopoietic stem cells and progenitors in liquid culture system. The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (119.5 +/- 5.7)% and (130.8 +/- 9.8)% respectively after 24 hours liquid culture (P < 0.05). The expansion effects of mBMEC-CM on CFU-GM and HPP-CFC were enhanced by compounded with flt3 ligand (FL) and thrombopoietin (TPO). The percentages of CFU-GM and HPP-CFC compared with the 0 hour control were (132.0 +/- 6.2)% and (176.9 +/- 12.8)% respectively after 24 hours liquid culture (P < 0.01).
Murine bone marrow endothelial cell conditioned medium could support the growth and proliferation of yolk sac hematopoitic stem cells and progenitors, and this promoting effect was further enhanced by addition of FL and TPO.
研究小鼠骨髓内皮细胞条件培养基（mBMEC-CM）对卵黄囊造血祖细胞生长的影响。
无血清mBMEC-CM取自于本实验室建立的源自骨髓的小鼠内皮细胞系的传代培养物。在交配后第8.5天收集小鼠卵黄囊，于37℃在含10%胎牛血清的0.1%胶原酶中孵育40分钟。卵黄囊细胞在组织培养皿中于37℃孵育1小时。非贴壁细胞在含10% mBMEC-CM和10%胎牛血清的DMEM中培养24小时后收集，用于粒细胞-巨噬细胞集落形成单位（CFU-GM）和高增殖潜能集落形成细胞（HPP-CFC）的半固体培养分析。分别在第7天和第14天计数CFU-GM和HPP-CFC的数量。
mBMEC-CM联合粒细胞巨噬细胞集落刺激因子（GM-CSF）可支持CFU-GM和HPP-CFC的生长。mBMEC-CM可在液体培养体系中诱导卵黄囊造血干细胞和祖细胞的增殖与分化。液体培养24小时后，CFU-GM和HPP-CFC相对于0小时对照组的百分比分别为（119.5±5.7）%和（130.8±9.8）%（P<0.05）。mBMEC-CM与fms样酪氨酸激酶3配体（FL）和血小板生成素（TPO）复合后对CFU-GM和HPP-CFC的扩增作用增强。液体培养24小时后，CFU-GM和HPP-CFC相对于0小时对照组的百分比分别为（132.0±6.2）%和（176.9±12.8）%（P<0.01）。
小鼠骨髓内皮细胞条件培养基可支持卵黄囊造血干细胞和祖细胞的生长与增殖，添加FL和TPO可进一步增强这种促进作用。
