Red blood cell lysis isolation and culture of rabbit bone marrow mesenchymal stem cells in vitro

doi:10.3969/j.issn.2095-4344.2013.49.002

Abstract

Abstract:

BACKGROUND: Some studies have demonstrated that red blood cell lysate added into the bone marrow can increase the efficiency of isolating and purifying mesenchymal stem cells, in order to obtain high-purity and high-quantity bone marrow mesenchymal stem cells.
OBJECTIVE: To isolate and culture bone marrow mesenchymal stem cells of rabbit with red blood cell lysis in vitro for exploring proliferation characteristics and performing the biological identification of cells.
METHODS: Bone marrow suspension was collected by puncturing the tibia with medullo-puncture needle. Red blood cell lysis was added to the bone marrow suspension, and bone marrow mesenchymal stem cells were isolated and cultured. Numbers of primary cells were recorded at 4, 7, 10, and 13 days later And Growth curves of the cells at passages 2-5 were drawn for comparison of proliferative characteristics. Inverted phase contrast microscope was used to observe the morphological changes of cells. CD34, CD44 antigens of bone marrow mesenchymal stem cells were identified by flow cytometry and immunity fluorescence.
RESULTS AND CONCLUSION: The adherent bone marrow mesenchymal stem cells mainly showed s spindle shape, with homogeneous nuclei, prominent nucleoli, and rich cytoplasm, which were positive for CD44 antigen and negative for CD34 antigen. The primary cells exhibited an “S” shape. Passage 4 cells had a better proliferative ability, rapider growth and more counting of cells as compared with other generations. These findings indicate that red blood cell lysis method is a feasible ways of culturing bone marrow mesenchymal stem cells 0 in vitro, and passage 4 cells have the strongest proliferation capacity.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: stem cells, mesenchymal stem cells, cells, cultured, cell proliferation

CLC Number:

R394.2

Zhang Wei-dong, Zhang Fang-biao, Shi Hong-can, Tan Rong-bang, Ye Gang, Li Guang-yu, Pan Shu, Sun Fei. Red blood cell lysis isolation and culture of rabbit bone marrow mesenchymal stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(49): 8468-8473.

Figures/Tables 5

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