兔骨髓间充质干细胞的分离培养与鉴定

Abstract

Abstract: Abstract: Objective To explore a safe and efficient method for isolation and identification of the rabbit bone marrow mesenchymal stem cells (BMSCs) in vitro,and to provide an experimental basis for the clinical application of BMSCs.Methods Bone marrow was aspirated from rabbit tibias,and the BMSCs were isolated and cultivated by total adherence method.The cells were observed by inverted phase contrast microscopes.The P5 cells were also induced to differentiate towards osteoblasts and chondroblasts,which were identified by corresponding histochemistry staining and FCM (flow cytometry).Results The primary and passage cells were spindleshaped and grew in colonies.ALP (alkaline phosphatase) activity increased continuously and the calcium nodule which was positive by Alizarin red staining was formed.The surface molecules including CD90,CD44 and CD105 were positive,and the cell surface markers including CD45,CD34 and Ckit were negative.Conclusions The adherence method is a simple and reliable method for rabbit BMSCs cultivation.BMSCs may be ideal seed cells for tissue engineering,which possess power of multidirectional differentiation.

Key words: mesenchymal stem cells, flow cytometry, tissue engineering.

CLC Number:

R329.29

References

[1] Friedenstein AJ, Gorskaja JF, Kulagina NN. Fibroblast precursors in normal and irradiated mouse hematopoietic organs [J]. Experimental Hematology, 1976, 4(5):267-274.
[2] 陈佳滨, 李强, 茹嘉, 等BMP-2和EGFP重组腺病毒体外转染兔骨髓间充质干细胞的研究[J]. 重庆医学, 2014, 43(2) :193-199
[3] Aldridge A, Kouroupis D, Churchman S, et al. Assay validation for the assessment of adipogenesis of multipotential stromal cells—a direct comparison of four different methods [J]. Cytotherapy, 2013, 15(1): 89-101.
[4] Cook D, Genever P. Regulation of mesenchymal stem cell differentiation [J]. Adv Exp Med Biol, 2013,786:213-29.
[5] Nandy S, Mohanty S, Singh M, et al. Fibroblast Growth Factor-2 alone as an efficient inducer for differentiation of human bone marrowmesenchymal stem cells into dopaminergic neurons[J]. J Biomed Sci, 2014, 21(1):83.
[6] Prockop DJ. Marrow stromal cells as stem cells for nonhematopoietic tissues [J]. Science, 1997, 276 (5309):71-74.
[7] Pittenger MF, Mackay AM, Beck SC, et al. Multilineage potential of adult human mesenchymal stem cells [J]. Science, 1999, 284:143-147.
[8] Amable PR, Teixeira MV, Carias RB, et al. Protein synthesis and secretion in human mesenchymal cells derived from bone marrow, adiposetissue and Wharton's jelly[J]. Stem Cell Res Ther, 2014, 16;5(2):53.
[9] Yanagisawa M, Yu RK. The expression and functions of glycoconjugates in neural stem cells [J]. Glycobiology, 2007, 17(7):57R-74R.
[10] Peister A, Mellad JA, Larson BL, et al. Adult stem cells from bone marrow (MSCs) isolated from different strains of inbred mice vary in surface epitopes, rates of proliferation, and differentiation potential [J]. Blood, 2004, 103(5):1662-1668.
[11] Xiang Z, Hu W, Kong Q, et al. Preliminary study of mesenchymal stem cells-seeded type I collagen-glycosaminoglycan matrices for cartilage repair[J]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi, 2006, 20(2):148-54.
[12] 章静波, 宗书东, 马文丽, 等. 干细胞[M]. 北京: 中国协和医科大学出版社, 2003, 1(10):130-135.
[13] Rasini V, Dominici M, Kluba T, et al. Mesenchymal stromal/stem cells markers in the human bone marrow [J]. Cytotherapy, 2013, 15(3): 292-306.
[14] Lian Q, Lye E, Suan Yeo K, et al. Derivation of clinically compliant MSCs from CD105+, CD24- differentiated human ESCs [J]. Stem Cells, 2007, 25(2):425-436.

[1]	. Effects of microspheres modified with collagen I on the adhesion, proliferation and osteogenesis of BMSCs [J]. , 2020, 40(4): 299-303.
[2]	. Osteogenic effects of small molecules on bone defect repair [J]. , 2019, 39(8): 739-743.
[3]	. Performance study of two new collagen membranes for guiding bone tissue regeneration in vivo and in vitro [J]. , 2019, 39(6): 481-487.
[4]	. Evaluation of Cytotoxicity of AlCoCrCuFeTix High Entropy Alloys and Its Effect on Apoptosis and Type I Collagen Synthesis in L929 Cells [J]. , 2019, 39(6): 500-504.
[5]	. Progress of mesenchymal stem cell related growth factor in alveolar bone repair [J]. , 2019, 39(1): 66-70.
[6]	. Effect of the growth differentiation factor 15 on rat bone marrowderived mesenchymal stem cell proliferation and gene expression [J]. , 2017, 37(9): 778-784.
[7]	. Study of the effect of platelet rich plasma on stem cell proliferation and differentiation [J]. , 2017, 37(8): 703-705.
[8]	. Effects of BMSCs and FG as seed cells and scaffold materials on repairing alveolar bone defect in rats [J]. , 2017, 37(4): 307-310.
[9]	FAN Zhipeng. Controlling the odontogenic differentiation of mesenchymal stem cells: the key to dental tissue regeneration [J]. , 2017, 37(2): 97-101.
[10]	. Research in the expression of LIM mineralization protein 1 during the mineralization process of two kinds of stem cells in vitro [J]. , 2017, 37(1): 20-24.
[11]	. Effects of hydrolic pressure on chondrogenic differentiation of bone marrow mesenchymal stem cells [J]. , 2016, 36(6): 481-484.
[12]	. A study on the odontogenic differentiation of placenta-derived mesenchymal stem cells in vitro [J]. , 2016, 36(5): 394-398.
[13]	. Autophagy in bone cells and its role in dental implantation [J]. , 2016, 36(5): 458-461.
[14]	. The effects of inflammatory microenvironment on regeneration of periodontal tissue [J]. , 2015, 35(9): 786-788.
[15]	. Features and function reconstruction of periodontal tissue: a key point for tooth regeneration in vivo [J]. , 2015, 35(8): 609-613.

Isolation and identification of the rabbit mesenchymal stem cells

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments