title: Enhanced viability and neural differential potential in poor post-thaw hADSCs by agarose multi-well dishes and spheroid culture.
abstract: Human adipose-derived stem cells (hADSCs) are potential adult stem cells source for cell therapy. But hADSCs with multi-passage or cryopreservation often revealed poor growth performance. The aim of our work was to improve the activity of poor post-thaw hADSCs by simple and effective means. We describe here a simple method based on commercially available silicone micro-wells for creating hADSCs spheroids to improve viability and neural differentiation potential on poor post-thaw hADSCs. The isolated hADSCs positively expresse d CD29, CD44, CD105, and negatively expressed CD34, CD45, HLA-DR by flow cytometry. Meanwhile, they had adipogenic and osteogenic differentiation capacity. The post-thaw and post-spheroid hADSCs from poor growth status hADSCs showed a marked increase in cell proliferation by CKK-8 analysis, cell cycle analysis and Ki67/P27 quantitative polymerase chain reaction (qPCR) analysis. They also displayed an increase viability of anti-apoptosis by annexin v and propidium iodide assays and mitochondrial membrane potential assays. After 3 days of neural induction, the neural differentiation potential of post-thaw and post-spheroid hADSCs could be enhanced by qPCR analysis and western blotting analysis. These results suggested that the spheroid formation could improve the viability and neural differentiation potential of bad growth status hADSCs, which is conducive to ADSCs research and cell therapy.
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Xiaoling
authors.initials: X
authors.lastname: Guo
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China.
authors.forename: Shanyi
authors.initials: S
authors.lastname: Li
authors.affiliation: Department of Ophthalmology, Affiliated Anhui Provincial Hospital of Anhui Medical University, Hefei, China.
authors.forename: Qingshan
authors.initials: Q
authors.lastname: Ji
authors.affiliation: Department of Ophthalmology, The First Clinical Medical College of Jinan University, Jinan University, Guangzhou, 510632, China.
authors.forename: Ruiling
authors.initials: R
authors.lastname: Lian
authors.affiliation: Key Laboratory for Regenerative Medicine, Ministry of Education, Jinan University, Guangzhou, China. chenjiansu2000@163.com.|Eye Institute, Medical College of Jinan University, Jinan University, Guangzhou, China. chenjiansu2000@163.com.|Department of Ophthalmology, The First Clinical Medical College of Jinan University, Jinan University, Guangzhou, 510632, China. chenjiansu2000@163.com.
authors.forename: Jiansu
authors.initials: J
authors.lastname: Chen
mesh_terms: D050153:Abdominal Fat / Q000166:cytology*; D015703:Antigens, CD; D018929:Cell Culture Techniques / Q000379:methods*; D002453:Cell Cycle; D002454:Cell Differentiation; D002470:Cell Survival; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D015925:Cryopreservation; D005260:Female; D006801:Humans; D019394:Ki-67 Antigen; D008875:Middle Aged; D009474:Neurons / Q000166:cytology*; D018809:Proliferating Cell Nuclear Antigen; D060888:Real-Time Polymerase Chain Reaction; D018874:Spheroids, Cellular; D013234:Stem Cells / Q000166:cytology*
chemical_list: D015703:Antigens, CD; D019394:Ki-67 Antigen; D018809:Proliferating Cell Nuclear Antigen; C413374:p27 antigen
keywords: Differentiation; Human adipose derived stem cells (hADSCs); Post-thaw hADSCs; Spheroid formation; Viability