Proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood under the culture conditions of hypoxia and serum deprivation

• Published in: Chinese medical journal Vol. 124; no. 23; pp. 3959 - 3967
• Main Authors: Fu, Wei-Li, Jia, Zhu-Qing, Wang, Wei-Ping, Zhang, Ji-Ying, Fu, Xin, Duan, Xiao-Ning, Leung, Kevin Kar Ming, Zhou, Chun-Yan, Yu, Jia-Kuo
• Format: Journal Article
• Language: English
• Published: China Institute of Sports Medicine, Peking University Third Hospital,Beijing 100191, China%Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Peking University Health Science Center,Beijing 100191, China 05.12.2011
• Subjects: Animals; Apoptosis - physiology; Cell Hypoxia - physiology; Cell Proliferation; Cells, Cultured; In Situ Nick-End Labeling; Mesenchymal Stromal Cells - cytology; Rabbits; 培养条件; 外周血; 细胞凋亡; 细胞增殖; 缺氧; 胎牛血清; 财产; 骨髓间充质干细胞; hypoxia; apoptosis; serum deprivation; proliferation; mesenchymal stem cells derived from peripheral blood
• ISSN: 0366-6999; 2542-5641
• DOI: 10.3760/cma.j.issn.0366-6999.2011.23.022

Background The proliferation and apoptosis property of mesenchymal stem cells derived from peripheral blood （PB-MSCs） were investigated under hypoxia and serum deprivation conditions in vitro so as to evaluate the feasibility for autologous PB-MSCs applications in cartilage repair. Methods MSCs were mobilized into peripheral blood by granulocyte colony stimulating factor （G-CSF） and AMD3100. The blood samples were collected from central ear artery of rabbits. Adhered cells were obtained by erythrocyte lysis buffer and identified as MSCs by adherence to plastic, spindle shaped morphology, specific surface markers, differentiation abilities into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. MSCs were cultured in four groups at different oxygen tension （20% 02 and 2% O2）, with or without 10% fetal bovine serum （FBS） conditions： 20% 02 and 10% FBS complete medium （normal medium, N）, 20% 02 and serum deprivation medium （D）, 2% 02 and 10% FBS complete medium （hypoxia, H）, 2% 02 and serum deprivation （HD）. Cell proliferation was determined by CCK-8 assay. Apoptosis was detected by Annexin V/PI and terminal deoxynucleotide transferase dUTP nick end labeling （TUNEL） staining. Results Spindle-shaped adherent cells were effectively mobilized from peripheral blood by a combined administration of G-CSF plus AMD3100. These cells showed typical fibroblast-like phenotype similar to MSCs from bone marrow （BM-MSCs）, and expressed a high level of typical MSCs markers CD29 and CD44, but lacked in the expression of hematopoietic markers CD45 and major histocompatibility complex Class II （MHC II）. They could also differentiate into osteoblasts, adipocytes and chondroblasts in vitro under appropriate conditions. No significant morphological differences were found among the four groups. It was found that hypoxia could enhance proliferation of PB-MSCs regardless of serum concentration, but serum deprivation inhibited proliferation at the later stage of culture. Apart from that, hypoxia or serum deprivation could promote the apoptosis of PB-MSCs after 48 hours; the effect was stronger when these two conditions combined together. Furthermore, the effect of serum deprivation on apoptosis was stronger compared with that of hypoxia. Conclusions PB-MSCs possess similar phenotypes as BM-MSCs. Their differentiation and proliferation abilities make them a new source of seed cells for ischemia-related cell therapy and tissue engineering in the field of the articular cartilage repair.