One-Step Derivation of Mesenchymal Stem Cell (MSC)-Like Cells from Human Pluripotent Stem Cells on a Fibrillar Collagen Coating

被引:170
作者
Liu, Yongxing [1 ]
Goldberg, A. Jon [1 ]
Dennis, James E. [2 ]
Gronowicz, Gloria A. [3 ]
Kuhn, Liisa T. [1 ]
机构
[1] Univ Connecticut, Ctr Hlth, Ctr Biomat, Dept Reconstruct Sci, Farmington, CT USA
[2] Benaroya Res Inst Virginia Mason, Seattle, WA USA
[3] Univ Connecticut, Ctr Hlth, Dept Surg, Farmington, CT USA
关键词
OSTEOGENIC DIFFERENTIATION; EX-VIVO; OSTEOBLAST PHENOTYPE; GENE-EXPRESSION; THIN-FILMS; GROWTH; CULTURE; SURVIVAL; ADHESION;
D O I
10.1371/journal.pone.0033225
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
Controlled differentiation of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) into cells that resemble adult mesenchymal stem cells (MSCs) is an attractive approach to obtain a readily available source of progenitor cells for tissue engineering. The present study reports a new method to rapidly derive MSC-like cells from hESCs and hiPSCs, in one step, based on culturing the cells on thin, fibrillar, type I collagen coatings that mimic the structure of physiological collagen. Human H9 ESCs and HDFa-YK26 iPSCs were singly dissociated in the presence of ROCK inhibitor Y-27632, plated onto fibrillar collagen coated plates and cultured in alpha minimum essential medium (alpha-MEM) supplemented with 10% fetal bovine serum, 50 uM magnesium L-ascorbic acid phosphate and 100 nM dexamethasone. While fewer cells attached on the collagen surface initially than standard tissue culture plastic, after culturing for 10 days, resilient colonies of homogenous spindle-shaped cells were obtained. Flow cytometric analysis showed that a high percentage of the derived cells expressed typical MSC surface markers including CD73, CD90, CD105, CD146 and CD166 and were negative as expected for hematopoietic markers CD34 and CD45. The MSC-like cells derived from pluripotent cells were successfully differentiated in vitro into three different lineages: osteogenic, chondrogenic, and adipogenic. Both H9 hES and YK26 iPS cells displayed similar morphological changes during the derivation process and yielded MSC-like cells with similar properties. In conclusion, this study demonstrates that bioimimetic, fibrillar, type I collagen coatings applied to cell culture plates can be used to guide a rapid, efficient derivation of MSC-like cells from both human ES and iPS cells.
引用
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页数:9
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