title: Feasibility of human hair follicle-derived mesenchymal stem cells/CultiSpher(®)-G constructs in regenerative medicine.
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.
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.|Department of Toxicology, School of Preventive Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Pengdong
authors.initials: P
authors.lastname: Li
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Feilin
authors.initials: F
authors.lastname: Liu
authors.affiliation: 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.
authors.forename: Chunling
authors.initials: C
authors.lastname: Wu
authors.affiliation: Department of Analytical Chemistry, School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Wenyue
authors.initials: W
authors.lastname: Jiang
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Guifang
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Tingting
authors.initials: T
authors.lastname: Bai
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Yixu
authors.initials: Y
authors.lastname: Jiang
authors.affiliation: Department of Pathology, Jilin Cancer Hospital, Changchun, Jilin, 130012, People's Republic of China.
authors.forename: Lili
authors.initials: L
authors.lastname: Guo
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Qi
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Junna
authors.initials: J
authors.lastname: Kou
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Ruirui
authors.initials: R
authors.lastname: Fan
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Deshun
authors.initials: D
authors.lastname: Hao
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Shaowei
authors.initials: S
authors.lastname: Lan
authors.affiliation: Department of Pathobiology, Key Laboratory of Ministry of Education, Norman Bethune College of Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China.
authors.forename: Yulin
authors.initials: Y
authors.lastname: Li
authors.affiliation: 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.|Department of Toxicology, School of Preventive Medicine, Jilin University, Changchun, Jilin, 130021, People's Republic of China. jy_liu@jlu.edu.cn.
authors.forename: Jin Yu
authors.initials: JY
authors.lastname: Liu
mesh_terms: D000328:Adult; D002454:Cell Differentiation; D049109:Cell Proliferation; D005260:Female; D018859:Hair Follicle / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D008875:Middle Aged; D044968:Regenerative Medicine
keywords: 3D cell culture; CultiSpher®-G microbeads; Hair follicle mesenchymal stem cells; Long-term expansion; Spinner culture