Cargando…
Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application
BACKGROUND: Pluripotent stem cell (PSC)-derived dopaminergic (DA) neurons are an expected source of cell therapy for Parkinson’s disease. The transplantation of cell aggregates or neurospheres, instead of a single cell suspension has several advantages, such as keeping the 3D structure of the donor...
Autores principales: | , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
IOS Press
2022
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9108593/ https://www.ncbi.nlm.nih.gov/pubmed/34958047 http://dx.doi.org/10.3233/JPD-212934 |
_version_ | 1784708740013883392 |
---|---|
author | Hiramatsu, Satoe Morizane, Asuka Kikuchi, Tetsuhiro Doi, Daisuke Yoshida, Kenji Takahashi, Jun |
author_facet | Hiramatsu, Satoe Morizane, Asuka Kikuchi, Tetsuhiro Doi, Daisuke Yoshida, Kenji Takahashi, Jun |
author_sort | Hiramatsu, Satoe |
collection | PubMed |
description | BACKGROUND: Pluripotent stem cell (PSC)-derived dopaminergic (DA) neurons are an expected source of cell therapy for Parkinson’s disease. The transplantation of cell aggregates or neurospheres, instead of a single cell suspension has several advantages, such as keeping the 3D structure of the donor cells and ease of handling. For this PSC-based therapy to become a widely available treatment, cryopreservation of the final product is critical in the manufacturing process. However, cryopreserving cell aggregates is more complicated than cryopreserving single cell suspensions. Previous studies showed poor survival of the DA neurons after the transplantation of cryopreserved fetal ventral-mesencephalic tissues. OBJECTIVE: To achieve the cryopreservation of induced pluripotent stem cell (iPSC)-derived DA neurospheres toward clinical application. METHODS: We cryopreserved iPSC-derived DA neurospheres in various clinically applicable cryopreservation media and freezing protocols and assessed viability and neurite extension. We evaluated the population and neuronal function of cryopreserved cells by the selected method in vitro. We also injected the cells into 6-hydroxydopamine (6-OHDA) lesioned rats, and assessed their survival, maturation and function in vivo. RESULTS: The iPSC-derived DA neurospheres cryopreserved by Proton Freezer in the cryopreservation medium Bambanker hRM (BBK) showed favorable viability after thawing and had equivalent expression of DA-specific markers, dopamine secretion, and electrophysiological activity as fresh spheres. When transplanted into 6-OHDA-lesioned rats, the cryopreserved cells survived and differentiated into mature DA neurons, resulting in improved abnormal rotational behavior. CONCLUSION: These results show that the combination of BBK and Proton Freezer is suitable for the cryopreservation of iPSC-derived DA neurospheres. |
format | Online Article Text |
id | pubmed-9108593 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | IOS Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-91085932022-05-18 Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application Hiramatsu, Satoe Morizane, Asuka Kikuchi, Tetsuhiro Doi, Daisuke Yoshida, Kenji Takahashi, Jun J Parkinsons Dis Research Report BACKGROUND: Pluripotent stem cell (PSC)-derived dopaminergic (DA) neurons are an expected source of cell therapy for Parkinson’s disease. The transplantation of cell aggregates or neurospheres, instead of a single cell suspension has several advantages, such as keeping the 3D structure of the donor cells and ease of handling. For this PSC-based therapy to become a widely available treatment, cryopreservation of the final product is critical in the manufacturing process. However, cryopreserving cell aggregates is more complicated than cryopreserving single cell suspensions. Previous studies showed poor survival of the DA neurons after the transplantation of cryopreserved fetal ventral-mesencephalic tissues. OBJECTIVE: To achieve the cryopreservation of induced pluripotent stem cell (iPSC)-derived DA neurospheres toward clinical application. METHODS: We cryopreserved iPSC-derived DA neurospheres in various clinically applicable cryopreservation media and freezing protocols and assessed viability and neurite extension. We evaluated the population and neuronal function of cryopreserved cells by the selected method in vitro. We also injected the cells into 6-hydroxydopamine (6-OHDA) lesioned rats, and assessed their survival, maturation and function in vivo. RESULTS: The iPSC-derived DA neurospheres cryopreserved by Proton Freezer in the cryopreservation medium Bambanker hRM (BBK) showed favorable viability after thawing and had equivalent expression of DA-specific markers, dopamine secretion, and electrophysiological activity as fresh spheres. When transplanted into 6-OHDA-lesioned rats, the cryopreserved cells survived and differentiated into mature DA neurons, resulting in improved abnormal rotational behavior. CONCLUSION: These results show that the combination of BBK and Proton Freezer is suitable for the cryopreservation of iPSC-derived DA neurospheres. IOS Press 2022-04-05 /pmc/articles/PMC9108593/ /pubmed/34958047 http://dx.doi.org/10.3233/JPD-212934 Text en © 2022 – The authors. Published by IOS Press https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY 4.0) License (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Research Report Hiramatsu, Satoe Morizane, Asuka Kikuchi, Tetsuhiro Doi, Daisuke Yoshida, Kenji Takahashi, Jun Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application |
title | Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application |
title_full | Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application |
title_fullStr | Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application |
title_full_unstemmed | Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application |
title_short | Cryopreservation of Induced Pluripotent Stem Cell-Derived Dopaminergic Neurospheres for Clinical Application |
title_sort | cryopreservation of induced pluripotent stem cell-derived dopaminergic neurospheres for clinical application |
topic | Research Report |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9108593/ https://www.ncbi.nlm.nih.gov/pubmed/34958047 http://dx.doi.org/10.3233/JPD-212934 |
work_keys_str_mv | AT hiramatsusatoe cryopreservationofinducedpluripotentstemcellderiveddopaminergicneurospheresforclinicalapplication AT morizaneasuka cryopreservationofinducedpluripotentstemcellderiveddopaminergicneurospheresforclinicalapplication AT kikuchitetsuhiro cryopreservationofinducedpluripotentstemcellderiveddopaminergicneurospheresforclinicalapplication AT doidaisuke cryopreservationofinducedpluripotentstemcellderiveddopaminergicneurospheresforclinicalapplication AT yoshidakenji cryopreservationofinducedpluripotentstemcellderiveddopaminergicneurospheresforclinicalapplication AT takahashijun cryopreservationofinducedpluripotentstemcellderiveddopaminergicneurospheresforclinicalapplication |