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Cell Tissue Res. 2015 Oct;362(1):69-86. doi: 10.1007/s00441-015-2182-z. Epub 2015 May 7.

Feasibility of human hair follicle-derived mesenchymal stem cells/CultiSpher(®)-G constructs in regenerative medicine.

Author information

1
Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
2
Department of Toxicology, School of Preventive Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
3
Harbin Veterinary Research Institute, CAAS-Michigan State University Joint Laboratory of Innate Immunity, State Key Laboratory of Veterinary Biotechnology, Chinese Academy of Agricultural Sciences, Maduan Street 427, Nangang District, Harbin, 150001, People's Republic of China.
4
Department of Analytical Chemistry, School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
5
Department of Pathology, Jilin Cancer Hospital, Changchun, Jilin, 130012, People's Republic of China.
6
Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China. jy_liu@jlu.edu.cn.
7
Department of Toxicology, School of Preventive Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China. jy_liu@jlu.edu.cn.

Abstract

The use of human mesenchymal stem cells (hMSCs) in cell therapies has increased the demand for strategies that allow efficient cell scale-up. Preliminary data on the three-dimensional (3D) spinner culture describing the potential use of microcarriers for hMSCs culture scale-up have been reported. We exploited a rich source of autologous stem cells (human hair follicle) and demonstrated the robust in vitro long-term expansion of human hair follicle-derived mesenchymal stem cells (hHF-MSCs) by using CultiSpher(®)-G microcarriers. We analyzed the feasibility of 3D culture by using hHF-MSCs/CultiSpher(®)-G microcarrier constructs for its potential applicability in regenerative medicine by comparatively analyzing the performance of hHF-MSCs adhered to the CultiSpher(®)-G microspheres in 3D spinner culture and those grown on the gelatin-coated plastic dishes (2D culture), using various assays. We showed that the hHF-MSCs seeded at various densities quickly adhered to and proliferated well on the microspheres, thus generating at least hundreds of millions of hHF-MSCs on 1 g of CultiSpher(®)-G within 12 days. This resulted in a cumulative cell expansion of greater than 26-fold. Notably, the maximum and average proliferation rates in 3D culture were significantly greater than that of the 2D culture. However, the hHF-MSCs from both the cultures retained surface marker and nestin expression, proliferation capacity and differentiation potentials toward adipocytes, osteoblasts and smooth muscle cells and showed no significant differences as evidenced by Edu incorporation, cell cycle, colony formation, apoptosis, biochemical quantification and qPCR assays.

KEYWORDS:

3D cell culture; CultiSpher®-G microbeads; Hair follicle mesenchymal stem cells; Long-term expansion; Spinner culture

PMID:
25948482
DOI:
10.1007/s00441-015-2182-z
[Indexed for MEDLINE]

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