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骨髓内皮细胞无血清条件培养液对骨髓内皮细胞增殖的促进作用
引用本文:Zhou XY,Wang QR,Huang YH,Cheng LM,Tan MQ. 骨髓内皮细胞无血清条件培养液对骨髓内皮细胞增殖的促进作用[J]. 生理学报, 2005, 57(2): 199-204
作者姓名:Zhou XY  Wang QR  Huang YH  Cheng LM  Tan MQ
作者单位:中南大学湘雅医学院生理学系血液生理研究室,中南大学生殖与干细胞工程研究所,长沙,410078;中南大学湘雅医学院生理学系血液生理研究室,中南大学生殖与干细胞工程研究所,长沙,410078;中南大学湘雅医学院生理学系血液生理研究室,中南大学生殖与干细胞工程研究所,长沙,410078;中南大学湘雅医学院生理学系血液生理研究室,中南大学生殖与干细胞工程研究所,长沙,410078;中南大学湘雅医学院生理学系血液生理研究室,中南大学生殖与干细胞工程研究所,长沙,410078
基金项目:This work was supported by the National Natural Science Foundation of China (No. 39970092) National High TECH Research and Development Programme of China (No.2003AA205018)and the Key Project of Science and Technology Programme of Hunan Province.
摘    要:本文通过制备小鼠骨髓内皮细胞无血清条件培养液(serum-free murine bone marrow endothelial cell conditioned medium, mBMEC-CM),经超滤分为分子量>10 kDa组分和<10 kDa组分,分别观察mBMEC-CM原液及其组分以及外源性细胞因子对小鼠骨髓内皮细胞集落生成的影响。用Wright’S Giemsa染色计数内皮细胞集落及检测骨髓内皮细胞的vWF,通过[3H]- TdR掺入量,观察mBMEC-CM原液及其组分以及外源性细胞因子对小鼠骨髓内皮细胞增殖的影响,并用分子杂交方法检测内皮细胞表达的细胞因子,从几个方面来研究mBMEC-CM对骨髓内皮细胞增殖的作用。结果显示,骨髓内皮细胞vWF 检测阳性。mBMEC-CM原液及其分子量>10 kDa组分能刺激骨髓内皮细胞集落增殖,且能明显增加骨髓内皮细胞[3H]-TdR 掺入量;分子量<10 kDa组分对骨髓内皮细胞集落增殖无明显刺激作用,也不能增加骨髓内皮细胞[3H]-TdR掺入量。外源加入IL-6、IL-11、SCF、GM-CSF、VEGF、bFGF 6种细胞因子能明显刺激骨髓内皮细胞集落增殖,SCF、VEGF、bFGF能明显增加骨髓内皮细胞[3H]-TdR掺入量。Atlas array膜杂交实验显示骨髓内皮细胞内源性表达GM-CSF、SCF、MSP-1、endothelin-2、thymosin β10、connective tissue GF、PDGF-A chain、MIP-2α、PlGF、neutrophil activating protein ENA-78、INF-γ、IL-1、IL-6、IL-13、IL-11、inhibin-α等细胞因子的mRNA。上述结果提示,骨髓内皮细胞无血清条件培养液对骨髓内皮细胞增殖具有促进作用。

关 键 词:骨髓内皮细胞  无血清条件培养液  增殖
修稿时间:2004-07-05

Promoting effects of serum-free murine bone marrow endothelial cell conditioned medium on the growth of bone marrow endothelial cells
Zhou Xiao-Ying,Wang Qi-Ru,Huang Yan-Hong,Cheng La-Mei,Tan Meng-Qun. Promoting effects of serum-free murine bone marrow endothelial cell conditioned medium on the growth of bone marrow endothelial cells[J]. Acta Physiologica Sinica, 2005, 57(2): 199-204
Authors:Zhou Xiao-Ying  Wang Qi-Ru  Huang Yan-Hong  Cheng La-Mei  Tan Meng-Qun
Affiliation:Experimental Hematology Laboratory, Department of Physiology, Xiangya School of Medicine; Reproduction and Stem Cell Engineering Institute, Central South University, Changsha 410078,China; E-mail: mqtan26@xysm.net.
Abstract:To study the effects of serum-free murine bone marrow endothelial cell conditioned medium (mBMEC-CM) on the growth of bone marrow endothelial cells, mBMEC-CM was collected and ultrafiltrated by Centriprep-10. The retentate of mBMEC-CM [molecular weight (MW)>10 kDa] and the filtrate of mBMEC-CM (MW<10 kDa) were obtained. The effect of bone marrow conditioned media, their components and exogenous cytokines on the formation of endothelial cell colonies were observed. The effect of bone marrow conditioned media, their components and exogenous cytokines on the proliferation of murine bone marrow endothelial cells were determined by [(3)H]-thymidine incorporation. The method of hybridizing to the Atlas cDNA array was used to determine the expression of cytokine mRNAs in bone marrow endothelial cells. The results obtained are as follows: vWF was expressed in bone marrow endothelial cells. The original mBMEC-CM and MW>10 kDa component of mBMEC-CM promoted the proliferation of bone marrow endothelial cell colonies and increased [(3)H]-thymidine incorporation of bone marrow endothelial cells. The MW<10 kDa component did not affect the production of endothelial cell colonies and did not increase [(3)H]-thymidine incorporation of endothelial cells. Six cytokines (IL-6, IL-11, SCF, GM-CSF, VEGF, bFGF) promoted the proliferation of bone marrow endothelial cell colonies. VEGF, bFGF and SCF increased [(3)H]-thymidine incorporation of bone marrow endothelial cells. According to the results of the Atlas cDNA array, GM-CSF,TGF-beta,BMP-2, bFGF, SCF, endothelin-2, thymosin beta10, MSP-1, connective tissue GF, PDGF-A chain, MIP-2 alpha, PlGF, neutrophil activating protein ENA-78, INF-gamma, IL-1, IL-6, IL-13, IL-11, inhibin-alpha mRNAs were expressed in endothelial cells. These results suggest that murine bone marrow endothelial cell conditioned medium promotes the proliferation of bone marrow endothelial cells.
Keywords:bone marrow endothelial cells  serum-free conditioned medium  proliferation
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