Bone marrow mesenchymal stem cells cultured in vitro and chemokine receptor CXCR4 expression

doi:10.3969/j.issn.2095-4344.2013.19.003

Abstract

Abstract:

BACKGROUND: Experiments have demonstrated that the expression level of CXCR4 on bone marrow mesenchymal stem cells determines cell homing to targeted tissue and subsequent angiogenesis.
OBJECTIVE: To analyze the isolation, culture, identification and multi-differentiation potential of bone marrow mesenchymal stem cells and investigate the significance of chemokine receptor CXCR4 expression on bone marrow mesenchymal stem cells in promotion of angiogenesis.
METHODS: Under sterile condition, bone marrow mesenchymal stem cells were isolated, cultured and purified by the while bone marrow adherence method. Cell apoptosis in vitro, CXCR4 protein expression on the bone marrow mesenchymal stem cells were detected, and cell cycle was analyzed.
RESULTS AND CONCLUSION: The isolated bone marrow mesenchymal stem cells exhibited uniform morphology. After passage, cells were spindle-shaped. Cells began to grow rapidly at about 4 days and grew stably 1 week later. Flow cytometry results showed increased CXCR4 expression on bone marrow mesenchymal stem cells. These findings suggest that bone marrow mesenchymal stem cells were successfully isolated, CXCR4 protein was highly expressed on cell surface after in vitro culture, and increased CXCR4 expression will provide theoretical evidence for bone marrow mesenchymal stem cell transplantation in promotion of angiogenesis and treatment of lower limb ischemic diseases in the clinic.

CLC Number:

R394.2

Yu Gui-ping, Shen Zhen-ya, Chen Guo-qiang, Yu Yun-sheng, Guo Shi-qiang. Bone marrow mesenchymal stem cells cultured in vitro and chemokine receptor CXCR4 expression[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(19): 3437-3442.

Figures/Tables 4

References

[1] Goldman BI, Wurzel J. Evidence that human cardiac myocytes divide after myocardial infarction. N Engl J Med. 2001;345(15):1131.

[2] Lange C,Brans H, Kluth D,et al.Hepatocytic differentiation of mesenchyrnal stem cells in eocuhures with fetai liver cells. World J Gastroenterol. 2006;12(15):2394-2397.

[3] Woedb D,Schwarz EJ,Prackop DJ,et al. Adutrat and human bone marrow stromal cells differentiate into neurons. J Neurosci Res. 2000;61(4):364-367.

[4] Krampera M,Marconi S,Pasini A,et al. Induction of neural-like differentiation in human mesenehymal stem cells derived from bone marrow, fat, spleen and thymus. Bone. 2007;40(2):382- 385.

[5] Cao Y, Lv G,Yu Y, et al. Liaoning Yixueyuan Xuebao. 2009; 30(2):105-106,115.曹阳,吕刚,于洋,等.趋化因子受体cxcr4在骨髓基质细胞中的表达及分析[J].辽宁医学院学报,2009,30(2):105-106,115.

[6] Zhang S, Jia Z, Ge J, et al. Purified human bone marrow multipotent mesenchymal stem cells regenerate infarcted myocardium in experimental rats. Cell Transplant. 2005; 14(10):787-798.

[7] Abarbanell AM, Coffey AC, Fehrenbacher JW, et al. Proinflammatory cytokine effects on mesenchymal stem cell therapy for the ischemic heart. Ann Thorac Surg. 2009;88(3): 1036-1043.

[8] Meirelles Lda S, Nardi NB. Murine marrow-derived mesenchymal stem cell: isolation, in vitro expansion, and characterization. Br J Haematol. 2003;123(4):702-711.

[9] Lian F, Zhu HS, Zhu W, et al. Isolation, culture and identification of porcine bone marrow mesenchymal stem cells and transformation to myogenic cells in vitro. Shanghai Laboratory Animal Science. 2003;23(2):67-69.

[10] Möhle R, Bautz F, Rafii S, et al. he chemokine receptor CXCR-4 is expressed on CD34+ hematopoietic progenitors and leukemic cells and mediates transendothelial migration induced by stromal cell-derived factor-1. Blood. 1998; 91(12):4523-4530.

[11] Cheng T. Stem cell fate outcomes modulated by the CDK inhibitors. Cell Research. 2008;18:s94.

[12] Kinnaird T, Stabile E, Burnett MS, et al. Marrow-derived stromal cells express genes encoding a broad spectrum of arteriogenic cytokines and promote in vitro and in vivo arteriogenesis through paracrine mechanisms. c Res. 2004; 94(5):678-685.

[13] Tropel P, Noël D, Platet N, et al. Isolation and characterisation of mesenchymal stem cells from adult mouse bone marrow. Exp Cell Res. 2004;295(2):395-406.

[14] Kitano Y, Radu A, Shaaban A, et al. Selection, enrichment, and culture expansion of murine mesenchymal progenitor cells by retroviral transduction of cycling adherent bone marrow cells. Exp Hematol. 2000;28(12):1460-1469.

[15] Tang YL, Zhao Q, Zhang YC, et al. Autologous mesenchymal stem cell transplantation induce VEGF and neovascularization in ischemic myocardium. Regul Pept. 2004;117(1):3-10.

[16] Tang J, Xie Q, Pan G, et al. Mesenchymal stem cells participate in angiogenesis and improve heart function in rat model of myocardial ischemia with reperfusion. Eur J Cardiothorac Surg. 2006;30(2):353-361.

[17] Tang J, Xie Q, Pan G, et al. Mesenchymal stem cells participate in angiogenesis and improve heart function in rat model of myocardial ischemia with reperfusion. Eur J Cardiothorac Surg. 2006;30(2):353-361.

[18] Zhu J, Zhou ZJ, Gong ZL, et al. Zhongguo Zuzhi Gongcheng Yanjiu yu Linchuang Kangfu. 2009;13(23):4485-4489.朱洁,周竹娟,龚自力,等.低氧对人骨髓间充质干细胞趋化因子受体CXCR4 和CX3CR1 表达的影响[J].中国组织工程研究与临床康复,2009,13(23):4485-4489.

[19] Pillarisetti K, Gupta SK. Cloning and relative expression analysis of rat stromal cell derived factor-1 (SDF-1)1: SDF-1 alpha mRNA is selectively induced in rat model of myocardial infarction. Inflammation. 2001;25(5):293-300.

[20] Peled A, Kollet O, Ponomaryov T, et al. The chemokine SDF-1 activates the integrins LFA-1, VLA-4, and VLA-5 on immature human CD34(+) cells: role in transendothelial/stromal migration and engraftment of NOD/SCID mice. Blood. 2000;95(11):3289-3296.

[21] Askari AT, Unzek S, Popovic ZB, et al. Effect of stromal-cell-derived factor 1 on stem-cell homing and tissue regeneration in ischaemic cardiomyopathy. Lancet. 2003;362 (9385):697-703.

[22] Clavin NW, Fernandez J, Schönmeyr BH, et al. Fractionated doses of ionizing radiation confer protection to mesenchymal stem cell pluripotency. Plast Reconstr Surg. 2008;122(3): 739-748.

[23] Zhu ZL, Mi N, Li J, et al. Jujie Shoushuxue Zazhi. 2010;19(6): 462-464.朱志立,米娜,李靖,等.趋化因子受体CXCR4在骨髓间充质干细胞中的表达[J].局解手术学杂志,2010,19(6):462-464.