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Promoting effects of serum--free murine bone marrow endothelial cell conditioned medium on the growth of bone marrow endothelial cells

Zhou Xiaoying, Wang Yiru, Huang Yanhong, Cheng Lamei, Tan Mengqun

Experimental Hematology Laboratory, Department of Physiology, Xiangya School of Medicine；China

Abstract

To study the effects of murine bone marrow endothelial cell conditioned medium on the growth of bone marrow endothelial cells, serum-free murine bone marrow endothelial cell conditioned medium (mBMEC-CM) was collected and ultrafiltration was performed with a centriprep 10 cut off molecular weight 10kD. The retentate of mBMEC-CM (mw>10kD) and the filtrate of mBMEC-CM (mw<10kD) 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>10kD 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<10kD component did not affect the production of endothelial cell colonies 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#beta#10, MSP-1, connective tissue GF, PDGF-A Chain, MIP-2#alpha#，PlGF，neutrophil activating protein ENA-78，INF-r，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 promoted the proliferation of bone marrow endothelial cells.

Key words: Bone marrow endothelial cells;serum-free condition medium;Proliferation

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Citing This Article：

Zhou Xiaoying, Wang Yiru, Huang Yanhong, Cheng Lamei, Tan Mengqun. Promoting effects of serum--free murine bone marrow endothelial cell conditioned medium on the growth of bone marrow endothelial cells. Acta Physiol Sin 2005; 57 (2): (in Chinese with English abstract).