Chinese Journal of Clinical Anatomy >
Study on biological characterization of Wharton's jelly mesenchymal stem cells from human umbilical cord
Received date: 2013-10-20
Online published: 2014-08-07
Objective To establish the culture methods of Wharton’s jelly mesenchymal stem cells from human umbilical cord（hWJ-MSCs）and observe the biological characteristics. Methods The umbilical cord organization of healthy full-term caesarean section was collected under sterile condition. The WJ was diced into cubes of approximately 0.5 mm3, and then plant them in culture flasks which kept still in 3 h before adding L-DMEM with 100 ml/L fetal bovine serum. The WJ-MSCs were passaged using 1.25 g/L Trypsin when they reached 90% confluence. Vimentin was tested by immunocytochemistry. The cell phenotype of WJ-MSCs was detected by flow cytometry. The hWJ-MSCs can differentiate into adipocytes like cell identified by oil-red stain. WJ-MSCs were cultured in medium containing 0.05 g//L Astragalus, then tested the expression of GFAP, NF, Nestin and NSE by immunocytochemical staining. Results Some cells swam out from the tissue block for 6 days or so, which belong to fibroblast-like cell appearing long spindle shaped fibroblast-like cells. WJ-MSCs after subculture presented in a uniform spindle morphology shape, showing parallel growth or vortex growth. The marks of MSCs such as CD90 were positive. The expression of CD34 was negative. Immunocytochemical staining showed WJ-MSCs expressed Vimentin with strong positive. After adipocyte like cells committed induction, these cells with lipid vacuoles accumulated in the cytoplasm gradually could be stained by oil-red O. After committed induction in 24 h by Astragalus, the differentiated cells were stained by Nestin and NF, but GFAP and NSE were negative. Conclusions WJ-MSCs can be effectively obtained from Wharton's jelly with the planting culture, which possess multi-differentiated capability.
Key words： Mesenchymal stem cells; Umbilical cord; Wharton's jelly; Isolation; Culture
LI Shi-Xin , HUA Yang-Kun , WANG Lin-Ping , YANG Hui , LU Dun , LV Chao-Chao , DONG Xu-Dong , SA E-Lian . Study on biological characterization of Wharton's jelly mesenchymal stem cells from human umbilical cord[J]. Chinese Journal of Clinical Anatomy, 2014 , 32(4) : 432 -436 . DOI: 10.13418/j.issn.1001-165x.2014.04.015
[1] Batsali AK, Kastrinaki MC, Papadaki HA, et al. Mesenchymal stem cells derived from Wharton's Jelly of the umbilical cord: biological properties and emerging clinical applications
[J]. Curr Stem Cell Res Ther, 2013, 8(2):144-155.
[2] 马云胜, 罗美, 刘树辉, 等. 人脐血基质细胞生物学特性的实验研究
[J].中国临床解剖学杂志, 2009, 27(2):216-219.
[3] Jin HJ, Bae YK, Kim M, et al. Comparative analysis of human mesenchymal stem cells from bone marrow, adipose tissue, and umbilical cord blood as sources of cell therapy
[J]. Int J Mol Sci, 2013, 14(9):17986-8001.
[4] Rao MS, Mattson MP. Stem cells and aging: expanding the possibilities
[J]. Mech Ageing Dev, 2001,122(6):713-734．
[5] Troyer DL, Weiss ML. Wharton's jelly-derived cells are a primitive stromal cell population
[J]. Stem Cells, 2008, 26(3):591-599.
[6] 董立华, 王勇, 陆长青. 黄芪诱导大鼠骨髓间充质干细胞分化为神经样细胞的研究
[J]. 四川大学学报（医学版）, 2007, 38(3): 417-420.
[7] Sarugaser R, Lickorish D, Baksh D, et al. Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors
[J]. Stem cells, 2005, 23(2): 220-229.
[8] Li WW, Wei YH, Li H, et al. Isolation and characterization of a novel strain of mesenchymal stem cells from mouse umbilical cord: potential application in cell-based therapy
[J]. PLoS One, 2013, 8(8):e74478.
[9] 高勇, 蒋明德, 王钊, 等. 一种分离培养人脐带Wharton's jelly间充质干细胞的新方法
[J]. 中国生物制品学杂志, 2013,26(6): 869-872.
[10] 庞荣清, 何洁, 李福兵, 等. 一种简单的人脐带间充质干细胞分离培养方法
[J]. 中华细胞与干细胞杂志（电子版）, 2011,1(2): 30-33.
[11] 撒亚莲, 华映坤, 高建梅，等. 人羊膜诱导人骨髓间充质干细胞向表皮样细胞的分化
[J]. 中国组织工程研究与临床康复, 2012,16(1):31-34.
[12]Emanuela P, Carlo C, Adele G, et al. Induction of dopaminergic neurons from human Wharton's jelly mesenchymal stem cell by forskolin
[J]. J Cell Physiol, 2014, 229(2):232-244.
[13]吴学建, 王军, 杜胜阳. 黄芪诱导人脐带间充质干细胞分化为神经样细胞的实验研究
[J]. 中华显微外科杂志, 2011, 34(2): 128-130.
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