Cryopreservation of undifferentiated and differentiated human neuronal cells
| Autor: | Kenji Yamatoya, Yuya Nagai, Naozumi Teramoto, Woojin Kang, Kenji Miyado, Kazuya Nakata, Tohru Yagi, Yoshitaka Miyamoto |
|---|---|
| Rok vydání: | 2022 |
| Předmět: |
Medicine (General)
SDS sodium dodecyl sulfate ESC embryonic stem cells Biomedical Engineering NeuN a neuronal marker DMSO dimethyl sulfoxide DAPI 4′ 6-Diamidino-2-phenylindole dihydrochloride D-PBS-free Dulbecco's phosphate-buffered saline without Ca2+ and Mg2+ supplementation Biomaterials iPSC induced pluripotent stem cells R5-920 FBS fetal bovine serum Sericin Cryopreservation QH573-671 Regenerative therapy D-MEM Dulbecco's Modified Eagle's Medium with high glucose l-glutamine phenol red and sodium pyruvate EDTA ethylene-diamine-tetraacetic acid 5-CFDA-AM 5-carboxyfluorescein diacetate Differentiation Neuronal cells Original Article BSA bovine serum albumin Cytology Serum-free Developmental Biology |
| Zdroj: | Regenerative Therapy Regenerative Therapy, Vol 19, Iss, Pp 58-68 (2022) |
| ISSN: | 2352-3204 |
| DOI: | 10.1016/j.reth.2021.12.007 |
| Popis: | The effective use of human-derived cells that are difficult to freeze, such as parenchymal cells and differentiated cells from stem cells, is crucial. A stable supply of damage-sensitive cells, such as differentiated neuronal cells, neurons, and glial cells can contribute considerably to cell therapy. We developed a serum-free freezing solution that is effective for the cryopreservation of differentiated neuronal cells. The quality of the differentiated and undifferentiated SK-N-SH cells was determined based on cell viability, live-cell recovery rate, and morphology of cultured cells, to assess the efficacy of the freezing solutions. The viability and recovery rate of the differentiated SK-N-SH neuronal cells were reduced by approximately 1.5-folds compared to that of the undifferentiated SK-N-SH cells. The viability and recovery rate of the differentiated SK-N-SH cells were remarkably different between the freezing solutions containing 10% DMSO and that containing 10% glycerol. Cryoprotectants such as fetal bovine serum (FBS), antifreeze proteins (sericin), and sugars (maltose), are essential for protecting against freeze damage in differentiated neuronal cells and parenchymal cells. Serum-free alternatives (sericin and maltose) could increase safety during cell transplantation and regenerative medicine. Considering these, we propose an effective freezing solution for the cryopreservation of neuronal cells. Graphical abstract Image 1 Highlights • The timing of freezing during cell differentiation. • More effective serum-free freezing solution for differentiated neuronal cells. • Improving the quality of damage-sensitive cells, such as differentiated neuronal cells. |
| Databáze: | OpenAIRE |
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