title: NASA-approved rotary bioreactor enhances proliferation of human epidermal stem cells and supports formation of 3D epidermis-like structure.
abstract: The skin is susceptible to different injuries and diseases. One major obstacle in skin tissue engineering is how to develop functional three-dimensional (3D) substitute for damaged skin. Previous studies have proved a 3D dynamic simulated microgravity (SMG) culture system as a "stimulatory" environment for the proliferation and differentiation of stem cells. Here, we employed the NASA-approved rotary bioreactor to investigate the proliferation and differentiation of human epidermal stem cells (hEpSCs). hEpSCs were isolated from children foreskins and enriched by collecting epidermal stem cell colonies. Cytodex-3 micro-carriers and hEpSCs were co-cultured in the rotary bioreactor and 6-well dish for 15 days. The result showed that hEpSCs cultured in rotary bioreactor exhibited enhanced proliferation and viability surpassing those cultured in static conditions. Additionally, immunostaining analysis confirmed higher percentage of ki67 positive cells in rotary bioreactor compared with the static culture. In contrast, comparing with static culture, cells in the rotary bioreactor displayed a low expression of involucrin at day 10. Histological analysis revealed that cells cultured in rotary bioreactor aggregated on the micro-carriers and formed multilayer 3D epidermis structures. In conclusion, our research suggests that NASA-approved rotary bioreactor can support the proliferation of hEpSCs and provide a strategy to form multilayer epidermis structure.
authors.affiliation: State Key Laboratory of Reproductive Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Xiao-hua
authors.initials: XH
authors.lastname: Lei
authors.forename: Li-na
authors.initials: LN
authors.lastname: Ning
authors.forename: Yu-jing
authors.initials: YJ
authors.lastname: Cao
authors.forename: Shuang
authors.initials: S
authors.lastname: Liu
authors.forename: Shou-bing
authors.initials: SB
authors.lastname: Zhang
authors.forename: Zhi-fang
authors.initials: ZF
authors.lastname: Qiu
authors.forename: Hui-min
authors.initials: HM
authors.lastname: Hu
authors.forename: Hui-shan
authors.initials: HS
authors.lastname: Zhang
authors.forename: Shu
authors.initials: S
authors.lastname: Liu
authors.forename: En-kui
authors.initials: EK
authors.lastname: Duan
mesh_terms: D019149:Bioreactors; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000502:physiology; D049109:Cell Proliferation; D000078404:Epidermal Cells; D004817:Epidermis / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006801:Humans; D019012:Integrin beta1 / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism
chemical_list: D019012:Integrin beta1
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