Tips and Tricks for Successfully Culturing and Adapting Human Induced Pluripotent Stem Cells [Protocol]
| Autor: | R. Castro-Viñuelas, Francisco J. Blanco, Isaac Fuentes-Boquete, Clara Sanjurjo-Rodríguez, Silvia Díaz-Prado, Isidoro López-Baltar, María Piñeiro-Ramil, Silvia Rodríguez-Fernández |
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| Rok vydání: | 2021 |
| Předmět: |
tips and tricks
Computer science Culture protocols iPSCs feeder-free Troubleshooting QH426-470 cryopreservation Regenerative medicine Tissue engineering Feeder-free Basic research Protocol Genetics Human Induced Pluripotent Stem Cells Induced pluripotent stem cell Molecular Biology Cryopreservation Feeders irradiation QH573-671 troubleshooting culture Transplantation Tips and tricks Molecular Medicine Irradiation feeders Cytology Neuroscience Reprogramming Trobleshooting Protocols |
| Zdroj: | RUC. Repositorio da Universidade da Coruña instname Molecular Therapy: Methods & Clinical Development, Vol 23, Iss, Pp 569-581 (2021) Molecular Therapy. Methods & Clinical Development |
| Popis: | Reprogramming somatic cells toward pluripotency became possible over a decade ago. Since then, induced pluripotent stem cells (iPSCs) have served as a versatile and powerful tool not only for basic research but also with the long-term goal of using them in human cell transplantation after differentiation. Nonetheless, downstream applications are frequently blurred by the difficulties that researchers have to face when working with iPSCs, such as trouble with clonal selection, in vitro culture and cryopreservation, adaptation to feeder-free conditions, or expansion of the cells. Therefore, in this article we aim to provide other researchers with practical and detailed information to successfully culture and adapt iPSCs. Specifically, we (1) describe the most common problems when in-vitro culturing iPSCs onto feeder cells as well as its possible troubleshooting, and (2) compare different matrices and culture media for adapting the iPSCs to feeder-free conditions. We believe that the troubleshooting and recommendations provided in this article can be of use to other researchers working with iPSCs and who may be experiencing similar issues, hopefully enhancing the appeal of this promising cell source to be used for biomedical investigations, such as tissue engineering or regenerative medicine applications. Graphical abstract This article contains a descriptive methodological guide on how to culture iPSCs under feeder or feeder-free conditions, paying special attention to provide readers with detailed protocols, tips and tricks, as well as troubleshooting guides than can be of use to other researchers working with iPSCs. |
| Databáze: | OpenAIRE |
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