Isolation, culture and identification of early and late endothelial progenitor cells from rat bone marrow and peripheral blood

doi:10.3969/j.issn.2095-4344.2013.06.018

Abstract

Abstract:

BACKGROUND: Our experiments intended to obtain bone marrow endothelial progenitor cells from rat peripheral blood and bone marrow, and culture the late endothelial progenitor cells to perform the stem cells transplantation or endothelial progenitor cells combined with gene therapy in order to highly express the angiogenic factors, thus to promote the angiogenesis after ischemic cerebrovascular disease.
OBJECTIVE: To isolate and identify the endothelial progenitor cells from rat bone marrow and peripheral blood.
METHODS: Mononuclear cells were obtained by using density gradient centrifugation and adherence screening method from bone marrow and peripheral blood of rats. Then the cells were induced, and the biological characteristics of the adherent cells were observed and recorded. Endothelial progenitor cells specific surface markers CD133, CD34 and KDR were selected and detected by immunofluorescence. Flow cytometry was used to detect the expression of KDR and CD34. Phagocytosis experiment was carried out to further identify the cells.
RESULTS AND CONCLUSION: Endothelial progenitor cells can be isolated from bone marrow and peripheral blood of rats; attached cells were positive for CD34, KDR and CD133 through immunofluorescence; cells were positive for CD34 and KDR by flow cytometry; cells could swallow acetylated-low-density lipoprotein, and bind with Ulex europaeus agglutinin 1. The endothelial progenitor cells were isolated from bone marrow and peripheral blood of rats successfully and the late endothelial progenitor cells that have strong proliferative activity were obtained. A better source of stem cells was found to form blood vessels.

Key words: stem cell, stem cell culture and differentiation, endothelial progenitor cells, bone marrow, peripheral blood, angiogenesis, cerebral ischemia stroke, stem cell photographs-containing paper

CLC Number:

R394.2

Wang Hong-juan, Wang Juan, Li Nan, Gao Hang, Liu Kang-ding. Isolation, culture and identification of early and late endothelial progenitor cells from rat bone marrow and peripheral blood[J]. Chinese Journal of Tissue Engineering Research, 2013, 17(6): 1056-1063.

Figures/Tables 10

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