Cryopreserved clumps of mesenchymal stem cell/extracellular matrix complexes retain osteogenic capacity and induce bone regeneration
| Autor: | Mikihito Kajiya, Susumu Horikoshi, Tsuyoshi Fujita, Souta Motoike, Katsuhiro Takeda, Nao Komatsu, Tomoyuki Iwata, Kazuhisa Ouhara, Shinji Matsuda, Manabu Takewaki, Hidemi Kurihara |
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| Rok vydání: | 2018 |
| Předmět: |
Male
0301 basic medicine Cell Survival Medicine (miscellaneous) MSCs Mesenchymal Stem Cell Transplantation Stem cell marker Biochemistry Genetics and Molecular Biology (miscellaneous) Cryopreservation lcsh:Biochemistry Extracellular matrix 03 medical and health sciences 0302 clinical medicine Osteogenesis Spheroids Cellular Animals C-MSC lcsh:QD415-436 Collagenases Bone regeneration Cells Cultured lcsh:R5-920 Chemistry Research Mesenchymal stem cell Artificial scaffold free Mesenchymal Stem Cells Cell Biology Ascorbic acid Rats Inbred F344 Extracellular Matrix Rats Cell biology Transplantation 030104 developmental biology 030220 oncology & carcinogenesis Molecular Medicine Stem cell lcsh:Medicine (General) |
| Zdroj: | Stem Cell Research & Therapy, Vol 9, Iss 1, Pp 1-13 (2018) Stem Cell Research & Therapy |
| ISSN: | 1757-6512 |
| Popis: | Background Three-dimensional (3D) cultured clumps of mesenchymal stem cell (MSC)/extracellular matrix (ECM) complexes (C-MSCs) consist of cells and self-produced ECM. C-MSCs can regulate cellular functions in vitro and can be grafted into a defect site without an artificial scaffold to induce bone regeneration. Long-term cryopreservation of C-MSCs, which can enable them to serve as a ready-to-use cell preparation, may be helpful in developing beneficial cell therapy for bone regeneration. Therefore, the aim of this study was to investigate the effect of cryopreservation on C-MSCs. Methods MSCs isolated from rat femurs were cultured in growth medium supplemented with ascorbic acid. To obtain C-MSCs, confluent cells that had formed on the cellular sheet were scratched using a micropipette tip and were then torn off. The sheet was rolled to make a round clumps of cells. The C-MSCs were cryopreserved in cryomedium including 10% dimethyl sulfoxide. Results Cryopreserved C-MSCs retained their 3D structure and did not exhibit a decrease in cell viability. In addition, stem cell marker expression levels and the osteogenic differentiation properties of C-MSCs were not reduced by cryopreservation. However, C-MSCs pretreated with collagenase before cryopreservation showed a lower level of type I collagen and could not retain their 3D structure, and their rates of cell death increased during cryopreservation. Both C-MSC and cryopreserved C-MSC transplantation into rat calvarial defects induced successful bone regeneration. Conclusion These data indicate that cryopreservation does not reduce the biological properties of C-MSCs because of its abundant type I collagen. More specifically, cryopreserved C-MSCs could be applicable for novel bone regenerative therapies. |
| Databáze: | OpenAIRE |
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