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Stemness and transdifferentiation of adipose-derived stem cells using L-ascorbic acid 2-phosphate-induced cell sheet formation.
Biomaterials, Issue: 11, Volume: 35, Pages: 3516-3526. | 2014-01-04
1 Patent citations    36 Scholarly citations     Reference Count: 47
Jiashing Yu;Yuan-Kun Tu;Yueh-Bih Tang;Nai-Chen Cheng

摘要

Cell sheet technology has emerged as an important tissue engineering approach. Adipose-derived stem cells (ASCs) have valuable applications in regenerative medicine, but their stemness and differentiation capabilities in the cell sheet format have not been well investigated. In this study, we found that L- ascorbate 2-phosphate (A2-P), a stable form of ascorbic acid, significantly enhanced ASC proliferation and induced ASC sheet fabrication in 7 days with abundant extracellular matrix deposition. Importantly, A2-P treatment significantly enhanced expression of pluripotent markers Sox-2, Oct-4 and Nanog, but treating ASCs with antioxidants other than A2-P revealed no stemness enhancement. Moreover, ASC treatment with A2-P and a collagen synthesis inhibitor, L-2-azetidine carboxylic acid or cis-4-hydroxy-D-proline, significantly inhibited the A2-P-enhanced expression of stemness markers. These findings demonstrated that A2-P enhances stemness of ASCs through collagen synthesis and cell sheet formation. We also showed that A2-P-stimulated collagen synthesis in ASCs may be mediated through ERK1/2 pathway. By culturing the ASC sheets in proper induction media, ASC transdifferentiation capabilities into neuron and hepatocyte-like cells were significantly enhanced after cell sheet formation, while adipogenic and osteogenic differentiation capacities were still maintained. Using a murine model of healing-impaired cutaneous wound, faster wound healing was noted in the group that received ASC sheet treatment, and we observed significantly more engrafted ASCs with evidence of differentiation toward endothelial and epidermal lineages in the cutaneous wound tissue. Therefore, A2-P-mediated ASC sheet formation enhanced ASC stemness and transdifferentiation capabilities, thereby representing a promising approach for applications in regenerative medicine.


机构

National Taiwan University;Department of Chemical Engineering, College of Engineering, National Taiwan University, Taipei, Taiwan.I-Shou University;Department of Orthopedics, E-Da Hospital/I-Shou University, Kaohsiung, Taiwan.National Taiwan University;Department of Surgery, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan.National Taiwan University;Department of Surgery, National Taiwan University Hospital and College of Medicine, Taipei, Taiwan; Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan. Electronic address: nccheng@ntu.edu.tw.


文献类型:

journal article;research support, non-u.s. gov't;

出版商:

Elsevier BV

发表时间:

2014-01-04

基金信息

E-Da Hospital-National Taiwan University Hospital Joint Research Program()