Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics
IntroductionIn vitro, primary rat oligodendrocytes (OLs) are widely used for research on OL development, physiology, and pathophysiology in demyelinating diseases such as multiple sclerosis. Primary culture methods for OLs from rats have been developed and improved over time, but there are still mul...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Cellular Neuroscience |
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| author | Hanki Kim Hanki Kim Bum Jun Kim Bum Jun Kim Seungyon Koh Seungyon Koh Seungyon Koh Hyo Jin Cho Byung Gon Kim Byung Gon Kim Jun Young Choi Jun Young Choi |
| author_facet | Hanki Kim Hanki Kim Bum Jun Kim Bum Jun Kim Seungyon Koh Seungyon Koh Seungyon Koh Hyo Jin Cho Byung Gon Kim Byung Gon Kim Jun Young Choi Jun Young Choi |
| author_sort | Hanki Kim |
| collection | DOAJ |
| description | IntroductionIn vitro, primary rat oligodendrocytes (OLs) are widely used for research on OL development, physiology, and pathophysiology in demyelinating diseases such as multiple sclerosis. Primary culture methods for OLs from rats have been developed and improved over time, but there are still multiple aspects in which efficiency can be boosted.MethodsTo make use of excess oligodendrocyte progenitor cells (OPCs) from primary cultures, a cryopreservation process utilizing a commercially available serum-free cryopreservation medium was established to passage and freeze OPCs at −80°C for later use.ResultsCryopreserved OPCs stored for up to 6 months were viable, and retained their OL lineage purity of ~98%. While OPCs cryopreserved for 3–6 months showed a decrease in cell density after two days of proliferation, ~17% of cryopreserved OPCs maintained the potential for proliferation comparable to control OPCs that had not frozen. After induction of differentiation for four days, ~43% of both control and cryopreserved OPCs differentiated into mature OLs, and when differentiation was induced on aligned nanofibers mimicking axonal structure, myelin sheath-like structures indicative of in vitro myelination was observed in all experimental groups.ConclusionThe validation of cryopreserved primary OLs as a functionally robust in vitro model can help improve the efficiency of primary OL culture, expand its applications, and reduce the inevitable sacrifice of animals. |
| format | Article |
| id | doaj-art-4d03506a89c243ebaff4e8be918c29dc |
| institution | Kabale University |
| issn | 1662-5102 |
| language | English |
| publishDate | 2025-01-01 |
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| series | Frontiers in Cellular Neuroscience |
| spelling | doaj-art-4d03506a89c243ebaff4e8be918c29dc2025-01-13T06:10:59ZengFrontiers Media S.A.Frontiers in Cellular Neuroscience1662-51022025-01-011810.3389/fncel.2024.15209921520992Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristicsHanki Kim0Hanki Kim1Bum Jun Kim2Bum Jun Kim3Seungyon Koh4Seungyon Koh5Seungyon Koh6Hyo Jin Cho7Byung Gon Kim8Byung Gon Kim9Jun Young Choi10Jun Young Choi11Department of Brain Science, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Biomedical Sciences, Ajou University Graduate School of Medicine, Suwon, Republic of KoreaDepartment of Brain Science, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Biomedical Sciences, Ajou University Graduate School of Medicine, Suwon, Republic of KoreaDepartment of Brain Science, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Biomedical Sciences, Ajou University Graduate School of Medicine, Suwon, Republic of KoreaDepartment of Neurology, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Brain Science, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Brain Science, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Neurology, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Brain Science, Ajou University School of Medicine, Suwon, Republic of KoreaDepartment of Neurology, Ajou University School of Medicine, Suwon, Republic of KoreaIntroductionIn vitro, primary rat oligodendrocytes (OLs) are widely used for research on OL development, physiology, and pathophysiology in demyelinating diseases such as multiple sclerosis. Primary culture methods for OLs from rats have been developed and improved over time, but there are still multiple aspects in which efficiency can be boosted.MethodsTo make use of excess oligodendrocyte progenitor cells (OPCs) from primary cultures, a cryopreservation process utilizing a commercially available serum-free cryopreservation medium was established to passage and freeze OPCs at −80°C for later use.ResultsCryopreserved OPCs stored for up to 6 months were viable, and retained their OL lineage purity of ~98%. While OPCs cryopreserved for 3–6 months showed a decrease in cell density after two days of proliferation, ~17% of cryopreserved OPCs maintained the potential for proliferation comparable to control OPCs that had not frozen. After induction of differentiation for four days, ~43% of both control and cryopreserved OPCs differentiated into mature OLs, and when differentiation was induced on aligned nanofibers mimicking axonal structure, myelin sheath-like structures indicative of in vitro myelination was observed in all experimental groups.ConclusionThe validation of cryopreserved primary OLs as a functionally robust in vitro model can help improve the efficiency of primary OL culture, expand its applications, and reduce the inevitable sacrifice of animals.https://www.frontiersin.org/articles/10.3389/fncel.2024.1520992/fulloligodendrocyteoligodendrocyte progenitor cell (OPC)primary culturecryopreservationin vitro myelination |
| spellingShingle | Hanki Kim Hanki Kim Bum Jun Kim Bum Jun Kim Seungyon Koh Seungyon Koh Seungyon Koh Hyo Jin Cho Byung Gon Kim Byung Gon Kim Jun Young Choi Jun Young Choi Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics Frontiers in Cellular Neuroscience oligodendrocyte oligodendrocyte progenitor cell (OPC) primary culture cryopreservation in vitro myelination |
| title | Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics |
| title_full | Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics |
| title_fullStr | Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics |
| title_full_unstemmed | Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics |
| title_short | Cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics |
| title_sort | cryopreservation of primary neonatal rat oligodendrocytes and recapitulation of in vitro oligodendrocyte characteristics |
| topic | oligodendrocyte oligodendrocyte progenitor cell (OPC) primary culture cryopreservation in vitro myelination |
| url | https://www.frontiersin.org/articles/10.3389/fncel.2024.1520992/full |
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