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Implantation of Bone Marrow Stromal Cell Sheets Derived from Old Donors Supports Bone Tissue Formation

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Tissue Engineering and Regenerative Medicine Aims and scope

Abstract

The purpose of this study was to evaluate the osteogenesis ability of osteogenic matrix cell sheets (OMCS) derived from old donor cells. Bone marrow stromal cells (BMSC) were obtained from young (7-week-old) and old (1-year-old) Fischer344 rats donors and cultured with modified Eagle’s medium (MEM group) alone or containing dexamethasone (Dex; 10 nM) and ascorbic acid phosphate (AscP; 0.28 mM) (Dex/AscP group). We prepared four in vitro experimental groups: (1) young MEM, (2) young Dex/AscP, (3) old MEM and (4) old Dex/AscP. Cell proliferation and osteogenic marker mRNA expression levels were evaluated in vitro. To assess bone formation in vivo, the cells of each group were combined with beta tricalcium phosphate (TCP) disks followed by implantation in recipient rats. The in vitro study showed significant differences in the mRNA expression of osteocalcin, ALP, and BMP2 between MEM and Dex/AscP groups. Bone formation following implantation was observed upon histological analyses of all groups. TCP combined with OMCS (OMCS/TCP group) resulted in enhanced bone formation compared to that following combination with BMSC (BMSC/TCP). The osteocalcin content of the OMCS/TCP group 4 weeks after implantation was significantly higher than that in the BMSC/TCP construct for both young and old donors. The present study clearly indicated that OMCS could be generated from BMSCs of old as well as young donors using a mechanical retrieval method. Thus, through its usage of OMCS, this method may represent a potentially effective therapeutic option for cell-based therapy in elderly patients.

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Acknowledgements

We thank Dr. T. Ueha for conducting a part of animal experiments and Dr. K. Kawate for interpretation of results. We also thank F. Kunda and M. Matsumura (Nara Medical University Faculty of Medicine, Japan) for their technical assistance. This study was partially supported by a Grant-in-Aid for Young Scientists (B).

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Authors and Affiliations

  1. Department of Public Health, Health Management and Policy, Faculty of Medicine, Nara Medical University, Shijo 840, Kashihara, Nara, 634-8521, Japan

    Manabu Akahane & Tomoaki Imamura

  2. Department of Orthopedic Surgery, Nara Medical University, Shijo 840, Kashihara, Nara, 634-8522, Japan

    Takamasa Shimizu, Tsutomu Kira, Takuya Egawa, Akinori Okuda, Tadanobu Onishi & Yasuhito Tanaka

  3. Department of Arthroplasty and Regenerative Medicine, Nara Medical University, Shijo 840, Kashihara, Nara, 634-8522, Japan

    Yusuke Inagaki

Authors
  1. Manabu Akahane
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  2. Takamasa Shimizu
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  3. Yusuke Inagaki
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  4. Tsutomu Kira
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  5. Takuya Egawa
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  6. Akinori Okuda
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  7. Tadanobu Onishi
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  8. Tomoaki Imamura
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  9. Yasuhito Tanaka
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Contributions

MA designed this study and wrote the first draft of the manuscript. TS and TK conducted animal experiments. TE, AO, and TO conducted biological analyses. YI, TI, and YT jointly interpreted the results.

Corresponding author

Correspondence to Manabu Akahane.

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The authors have no conflict of interest to disclose.

Ethical statement

All experimental protocols using animals were approved by the Animal Experimental Review Board of Nara Medical University (IRB No. 11345) prior to initiating the experiments. The animals were housed in a temperature-controlled environment at approximately 21 °C under a 12 h light/12 h dark cycle with free access to food and water.

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Akahane, M., Shimizu, T., Inagaki, Y. et al. Implantation of Bone Marrow Stromal Cell Sheets Derived from Old Donors Supports Bone Tissue Formation. Tissue Eng Regen Med 15, 89–100 (2018). https://doi.org/10.1007/s13770-017-0088-4

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  • DOI: https://doi.org/10.1007/s13770-017-0088-4

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