Evaluation of different protocols for culturing mesenchymal stem cells derived from murine bone marrow
| Autor: | Mariana Lazarini, Cristiane Okuda Torello, Mariana Ferreira Pissarra, Sara Teresinha Olalla Saad |
|---|---|
| Přispěvatelé: | UNIVERSIDADE ESTADUAL DE CAMPINAS |
| Rok vydání: | 2022 |
| Předmět: |
Técnicas in vitro
Camundongo Medula óssea Flow cytometry Mice In vitro techniques Tissue culture Tissue engineering medicine Artigo original Immunology and Allergy Bone marrow Técnicas de cultura de células medicine.diagnostic_test Células mesenquimais estromais business.industry Mesenchymal stem cell Hematology Molecular biology Haematopoiesis medicine.anatomical_structure Cell culture Mesenchymal stem cells business Cell culture techniques Fetal bovine serum |
| Zdroj: | Repositório da Produção Científica e Intelectual da Unicamp Universidade Estadual de Campinas (UNICAMP) instacron:UNICAMP Repositório Institucional da Unicamp |
| ISSN: | 2531-1379 |
| DOI: | 10.1016/j.htct.2021.02.005 |
| Popis: | Agradecimentos: This study was supported by Sao Paulo Research Foundation (FAPESP - grants 2017/19674-2, 2017/21801-2), National Council for Scientific and Technological Development (CNPq) and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior Brasil (CAPES) - Finance Code 001 Abstract: Culturing bone marrow mesenchymal stem cells (BM-MSCs) is a key point in different fields of research, including tissue engineering and regenerative medicine and studies of the bone marrow microenvironment. However, isolating and expanding murine BM-MSCs in vitro has challenged researchers due to the low purity and yield of obtained cells. In this study, we aimed to evaluate five different protocols to culture murine BMMSCs in vitro. Methods: All protocols were based on the adhesion capacity of BM-MSCs to the tissue culture plastic surface and varied in the types of plate, culture media, serum, additional supplementation and initial cell density. Flow cytometry analysis was used to investigate lineage purity after expansion. Results: The expression of CD45 and CD11b was detected in the cultures generated according to all protocols, indicating low purity with the presence of hematopoietic cells and macrophages. The cellular growth rate and morphology varied between the cultures performed according to each protocol. Cells cultured according to protocol 5 (8 £ 107 cells/plate, Roswell Park Memorial Institute (RPMI) culture medium during first passage and then Iscove's Modified Delbecco's Medium (IMDM) culture medium, both supplemented with 9% fetal bovine serum, 9% horse serum, 12mM L-glutamine) presented the best performance, with a satisfactory growth rate and spindle-shape morphology. Conclusion: Our results point out that the purity and satisfactory growth rate of murine BMMSC cultures are not easily achieved and additional approaches must be tested for a proper cell expansion FUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO (FAPESP) CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO (CNPQ) COORDENAÇÃO DE APERFEIÇOAMENTO DE PESSOAL DE NÍVEL SUPERIOR (CAPES) Fechado |
| Databáze: | OpenAIRE |
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