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[Mechanism of granulocyte colony-stimulating factor for promoting cell viability of bone marrow mesenchymal stem cells.] [Ariticle in Chinese]

Chen Long, CHENG Fan-Jun, LIU Qi-Huan, TANG Jun-Ming, ZENG Qin-Bing, KONG Xia, GUO Ling-Yun, WANG Jia-Ning

1Department of Hematology, Dongfeng Hospital, Yunyang Medical College, Shiyan 442006, China； Institute of Clinical Medicine, Yunyang Medical College, Shiyan 442000, China

Abstract

The present study was aimed to investigate the mechanism of the granulocyte colony-stimulating factor (G-CSF) on the viability of the bone marrow mesenchymal stem cells (MSCs). MSCs were cultured by classical whole bone marrow adhering method, and the MSCs were analyzed for the cell surface differentiation markers CD34, CD133, CD90 and CD105 by flow cytometry (FCM). The ability of the MSCs to differentiate into osteocytes and adipocytes was tested in osteogenic and adipogenic mediums, separately. The effect of G-CSF (20 μg/mL) on the passage 3 MSCs viability was evaluated by MTT method, and the molecular mechanism of the G-CSF mediated effects was assayed through the pretreatment of the signal pathway inhibitors including 50 nmol/L wortmannin (phosphatidylinoesitol 3 kinase inhibitor), 50 μmol/L PD98059 [extracellular signal-regulated-kinase1/2 (ERK1/2) inhibitor], 30 μmol/L SB203580 (p38 mitogen-activated protein kinase inhibitor), 10 μmol/L H89 (protein kinase A inhibitor), 20 μmol/L Y27632 (Rho kinase inhibitor), 1 μmol/L rapamycin [mammalian target of rapamycin (mTOR) inhibitor], 10 mmol/L straurosporine [protein kinase C (PKC) inhibitor], 6 nmol/L G0697 (PKCα inhibitor) and 50 μmol/L Pseudo Z (PKCζ inhibitor). Cultured passage 3 MSCs expressed CD90 and CD105 strongly, and showed the ability of multi-differentiation into osteocytes and adipocytes. G-CSF promoted the viability of MSCs, and the promotion was completely inhibited by PKC inhibitor straurosporine and partially inhibited by wortmannin, rapamycin, PD98059, SB203580 or G0697. However, its effect was not inhibited by H89, Y27632 and Pseudo Z. It is thus suggested that the promoting effect of G-CSF on MSCs viability was closely related to AKT-mTOR-PKC signal pathway, and PKC maybe the central role in the signal pathway.

Key words: granulocyte colony-stimulating factor; mesenchymal stem cell; protein kinase C

Received: 2008-06-04　　Accepted: 2009-01-15

Corresponding author: 程范军　　E-mail: chengfanjun001@sina.com

Citing This Article：

Chen Long, CHENG Fan-Jun, LIU Qi-Huan, TANG Jun-Ming, ZENG Qin-Bing, KONG Xia, GUO Ling-Yun, WANG Jia-Ning. [Mechanism of granulocyte colony-stimulating factor for promoting cell viability of bone marrow mesenchymal stem cells.] [Ariticle in Chinese] . Acta Physiol Sin 2009; 61 (2): 169-174 (in Chinese with English abstract).