CryoPause: A New Method to Immediately Initiate Experiments after Cryopreservation of Pluripotent Stem Cells
| Autor: | Siera A. Rosen, Elisa de Stanchina, Chao Zhang, Karen G. Wong, Mark J. Tomishima, Franz-Josef Müller, Kiran Ramnarine, Sean D. Ryan, Amanda Kulick, Doron Betel, Thadeous J. Kacmarczyk, Shannon E. Mann |
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| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Resource Pluripotent Stem Cells Cellular differentiation Cell- and Tissue-Based Therapy Biology Biochemistry Cryopreservation Cell Line 03 medical and health sciences Mice 0302 clinical medicine Directed differentiation disease modeling Genetics human pluripotent stem cell cell banking Animals Humans quality control Induced pluripotent stem cell lcsh:QH301-705.5 Biological Specimen Banks Gene Editing lcsh:R5-920 business.industry directed differentiation Cell Differentiation Cell Biology Biotechnology 030104 developmental biology lcsh:Biology (General) Biochemical engineering cell therapy business lcsh:Medicine (General) 030217 neurology & neurosurgery Developmental Biology |
| Zdroj: | Stem Cell Reports Stem Cell Reports, Vol 9, Iss 1, Pp 355-365 (2017) |
| ISSN: | 2213-6711 |
| Popis: | Summary Human pluripotent stem cells (PSCs) provide an unlimited cell source for cell therapies and disease modeling. Despite their enormous power, technical aspects have hampered reproducibility. Here, we describe a modification of PSC workflows that eliminates a major variable for nearly all PSC experiments: the quality and quantity of the PSC starting material. Most labs continually passage PSCs and use small quantities after expansion, but the “just-in-time” nature of these experiments means that quality control rarely happens before use. Lack of quality control could compromise PSC quality, sterility, and genetic integrity, which creates a variable that might affect results. This method, called CryoPause, banks PSCs as single-use, cryopreserved vials that can be thawed and immediately used in experiments. Each CryoPause bank provides a consistent source of PSCs that can be pre-validated before use to reduce the possibility that high levels of spontaneous differentiation, contamination, or genetic integrity will compromise an experiment. Highlights • CryoPause is a new way to perform PSC-based experiments that enhances reproducibility • Large batches of single-use, cryopreserved cells are used directly post thaw • Directed differentiation and gene editing can be done without expansion • Repeated experiments can be done from the same PSCs over time and geographical place PSC work has traditionally been performed by passaging cells over time while retaining a portion of culture to perform work. Such extended culture creates confounding variables that can skew results. Tomishima and colleagues demonstrate that PSCs can be biobanked as “ready-to-use” aliquots so that PSCs can be thawed before immediate use in directed differentiation or gene editing experiments. |
| Databáze: | OpenAIRE |
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