Reproducible methodology for the isolation of mesenchymal stem cells from human umbilical cord and its potential for cardiomyocyte generation

被引:97
作者
Pereira, Winston Costa [1 ]
Khushnooma, Italia [2 ]
Madkaikar, Manisha [2 ]
Ghosh, K. [2 ]
机构
[1] LifeCell Int Private Ltd, R&D, Madras 600048, Tamil Nadu, India
[2] King Edward Mem Hosp, Inst Immunohaematol, Bombay, Maharashtra, India
关键词
mesenchymal stem cells; collagenase; umbilical cord; characterization; differentiation; cardiomyocytes;
D O I
10.1002/term.107
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Mesenchymal stem cells (MSCs) are considered to be a source of stem cells in tissue regeneration and therapeutics, due to their ability to undergo proliferation and differentiation. Complications associated with bone marrow-derived MSCs has prompted researchers to explore alternative sources of MSCs. The human umbilical cord is one such source; it is easily available and its collection is non-invasive. The sources of MSCs are non-controversial and thus they are not subjected to ethical constraints, as in the case of embryonic stem cells. MSCs are multipotent stem cells and has the ability to differentiate into various cell types of the mesodermal lineage. The aim of this study was to establish a reproducible method for the isolation of MSCs from human umbilical cord, as the few methods published till date gave inconsistent results and had a mixed population of contaminating endothelial cells. In our isolation strategy, we isolated a pure population of MSCs from Wharton's jelly of the human umbilical cord, which is very rich in collagen, and we used a high concentration of collagenase enzyme in the isolation of MSCs. Extensive phenotypic characterization analysis of these cells, using flow cytometry and antibody staining methods, have shown that we were able to isolate a pure population of the mesenchymal lineage cells that is devoid of haematopoietic and endothelial cell contaminants. When these MSCs were subjected to cardiomyocyte differentiation, we observed a change in the morphological characteristics, which was accompanied by the formation of myotube structures and spontaneous beating after 21 days. Copyright (C) 2008 John Wiley & Sons, Ltd.
引用
收藏
页码:394 / 399
页数:6
相关论文
共 25 条
[1]   Mesenchymal stem cells: building blocks for molecular medicine in the 21st century [J].
Caplan, AI ;
Bruder, SP .
TRENDS IN MOLECULAR MEDICINE, 2001, 7 (06) :259-264
[2]   Expression of early transcription factors Oct-4, Sox-2 and Nanog by porcine umbilical cord (PUC) matrix cells [J].
Carlin, R ;
Davis, D ;
Weiss, M ;
Schultz, B ;
Troyer, D .
REPRODUCTIVE BIOLOGY AND ENDOCRINOLOGY, 2006, 4 (1)
[3]   Antigen-presenting property of mesenchymal stem cells occurs during a narrow window at low levels of interferon-γ [J].
Chan, Jennifer L. ;
Tang, Katherine C. ;
Patel, Anoop P. ;
Bonilla, Larissa M. ;
Pierobon, Nicola ;
Ponzio, Nicholas M. ;
Rameshwar, Pranela .
BLOOD, 2006, 107 (12) :4817-4824
[4]   Detection of stromal cells in peripheral blood progenitor cell collections from breast cancer patients [J].
Fernandez, M ;
Simon, V ;
Herrera, G ;
Cao, C ;
DelFavero, H ;
Minguell, JJ .
BONE MARROW TRANSPLANTATION, 1997, 20 (04) :265-271
[5]   DEVELOPMENT OF FIBROBLAST COLONIES IN MONOLAYER CULTURES OF GUINEA-PIG BONE MARROW AND SPLEEN CELLS [J].
FRIEDENSTEIN, AJ ;
CHAILAKHJAN, RK ;
LALYKINA, KS .
CELL AND TISSUE KINETICS, 1970, 3 (04) :393-+
[6]  
Izadpanah Reza, 2008, V449, P153, DOI 10.1007/978-1-60327-169-1_11
[7]   Mesenchymal stromal cells [J].
Keating, Armand .
CURRENT OPINION IN HEMATOLOGY, 2006, 13 (06) :419-425
[8]   Discarded stem cells with a future? [J].
Klingemann, Hans .
EXPERT OPINION ON BIOLOGICAL THERAPY, 2006, 6 (12) :1251-1254
[9]   Isolation of multipotent mesenchymal stem cells from umbilical cord blood [J].
Lee, OK ;
Kuo, TK ;
Chen, WM ;
Lee, KD ;
Hsieh, SL ;
Chen, TH .
BLOOD, 2004, 103 (05) :1669-1675
[10]  
Lu LL, 2006, HAEMATOLOGICA, V91, P1017