Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells

Introduction: Stromal vascular fraction (SVF), a heterogeneous cell population primarily derived from adipose tissue, is widely utilized in regenerative therapies for its wound-healing properties and accessibility. While its immediate availability is advantageous, repeated harvesting can be burdenso...

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Main Authors: Naoki Inafuku, Yoshihiro Sowa, Tsunao Kishida, Seiji Sawai, Edward Hosea Ntege, Toshiaki Numajiri, Kenta Yamamoto, Yusuke Shimizu, Osam Mazda
Format: Article
Language:English
Published: Elsevier 2025-06-01
Series:Regenerative Therapy
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Online Access:http://www.sciencedirect.com/science/article/pii/S2352320425000306
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author Naoki Inafuku
Yoshihiro Sowa
Tsunao Kishida
Seiji Sawai
Edward Hosea Ntege
Toshiaki Numajiri
Kenta Yamamoto
Yusuke Shimizu
Osam Mazda
author_facet Naoki Inafuku
Yoshihiro Sowa
Tsunao Kishida
Seiji Sawai
Edward Hosea Ntege
Toshiaki Numajiri
Kenta Yamamoto
Yusuke Shimizu
Osam Mazda
author_sort Naoki Inafuku
collection DOAJ
description Introduction: Stromal vascular fraction (SVF), a heterogeneous cell population primarily derived from adipose tissue, is widely utilized in regenerative therapies for its wound-healing properties and accessibility. While its immediate availability is advantageous, repeated harvesting can be burdensome, especially for elderly patients, and the regenerative capacity of SVF declines with donor age. Long-term cryopreservation offers a potential solution by allowing the banking of SVF from younger donors for future use; however, the impact of this process on SVF functionality remains elusive. This study investigates the stemness and wound-healing potential of SVF following prolonged cryopreservation. Methods: SVF cells were isolated from adipose tissue harvested from twelve patients and cryopreserved for either two months (short-term cryopreserved SVF, S-SVF) or 12–13 years (long-term cryopreserved SVF, L-SVF), with six patients in each group. In vitro assays assessed cell viability and stemness, while in vivo assays evaluated wound-healing ability by administering thawed SVF cells from each group to dorsal wounds in immunodeficient mice, compared with a control group. Non-parametric statistical tests analyzed the differences between groups. Results: L-SVF exhibited significantly lower stemness compared to S-SVF. Importantly, the L-SVF group showed significantly improved wound healing compared with the control group, although the wound-healing effect of L-SVF was inferior to that of the S-SVF. Conclusion: This study demonstrated that, despite reduced stemness, L-SVF retains partial wound-healing potential after 12–13 years of cryopreservation. These findings highlight the need for optimized cryopreservation protocols to enhance SVF viability and regenerative capacity for clinical applications.
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spelling doaj-art-80c570faa54d401dbe06789fcf66e59c2025-08-20T01:51:16ZengElsevierRegenerative Therapy2352-32042025-06-012912813910.1016/j.reth.2025.02.004Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cellsNaoki Inafuku0Yoshihiro Sowa1Tsunao Kishida2Seiji Sawai3Edward Hosea Ntege4Toshiaki Numajiri5Kenta Yamamoto6Yusuke Shimizu7Osam Mazda8Department of Plastic and Reconstructive Surgery, Kyoto Prefectural University of Medicine, Kamigyo, Kyoto, JapanDepartment of Plastic Surgery, Jichi Medical University, Shimotsuke, Tochigi, Japan; Corresponding author.Department of Immunology, Kyoto Prefectural University of Medicine, Kamigyo, Kyoto, JapanDepartment of Orthopedics, Jyujyo Takeda Rehabilitation Hospital, Kyoto, JapanDepartment of Plastic and Reconstructive Surgery, Graduate School of Medicine, University of the Ryukyus, Okinawa, JapanDepartment of Plastic and Reconstructive Surgery, Kyoto Prefectural University of Medicine, Kamigyo, Kyoto, JapanDepartment of Immunology, Kyoto Prefectural University of Medicine, Kamigyo, Kyoto, JapanDepartment of Plastic and Reconstructive Surgery, Graduate School of Medicine, University of the Ryukyus, Okinawa, JapanDepartment of Immunology, Kyoto Prefectural University of Medicine, Kamigyo, Kyoto, JapanIntroduction: Stromal vascular fraction (SVF), a heterogeneous cell population primarily derived from adipose tissue, is widely utilized in regenerative therapies for its wound-healing properties and accessibility. While its immediate availability is advantageous, repeated harvesting can be burdensome, especially for elderly patients, and the regenerative capacity of SVF declines with donor age. Long-term cryopreservation offers a potential solution by allowing the banking of SVF from younger donors for future use; however, the impact of this process on SVF functionality remains elusive. This study investigates the stemness and wound-healing potential of SVF following prolonged cryopreservation. Methods: SVF cells were isolated from adipose tissue harvested from twelve patients and cryopreserved for either two months (short-term cryopreserved SVF, S-SVF) or 12–13 years (long-term cryopreserved SVF, L-SVF), with six patients in each group. In vitro assays assessed cell viability and stemness, while in vivo assays evaluated wound-healing ability by administering thawed SVF cells from each group to dorsal wounds in immunodeficient mice, compared with a control group. Non-parametric statistical tests analyzed the differences between groups. Results: L-SVF exhibited significantly lower stemness compared to S-SVF. Importantly, the L-SVF group showed significantly improved wound healing compared with the control group, although the wound-healing effect of L-SVF was inferior to that of the S-SVF. Conclusion: This study demonstrated that, despite reduced stemness, L-SVF retains partial wound-healing potential after 12–13 years of cryopreservation. These findings highlight the need for optimized cryopreservation protocols to enhance SVF viability and regenerative capacity for clinical applications.http://www.sciencedirect.com/science/article/pii/S2352320425000306CryopreservationStromal vascular fractionAdipose-derived stem cellsWound-healingRegenerative medicineLong-term preservation
spellingShingle Naoki Inafuku
Yoshihiro Sowa
Tsunao Kishida
Seiji Sawai
Edward Hosea Ntege
Toshiaki Numajiri
Kenta Yamamoto
Yusuke Shimizu
Osam Mazda
Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells
Regenerative Therapy
Cryopreservation
Stromal vascular fraction
Adipose-derived stem cells
Wound-healing
Regenerative medicine
Long-term preservation
title Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells
title_full Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells
title_fullStr Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells
title_full_unstemmed Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells
title_short Investigation of the stemness and wound-healing potential of long-term cryopreserved stromal vascular fraction cells
title_sort investigation of the stemness and wound healing potential of long term cryopreserved stromal vascular fraction cells
topic Cryopreservation
Stromal vascular fraction
Adipose-derived stem cells
Wound-healing
Regenerative medicine
Long-term preservation
url http://www.sciencedirect.com/science/article/pii/S2352320425000306
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