Skip to main content
SpringerLink
Log in

Isolation of adipose tissue-derived stem cells by direct membrane migration and expansion for clinical application

  • Research Article
  • Published:
Human Cell Aims and scope Submit manuscript

Abstract

Mesenchymal stem cells (MSCs) have recently made significant progression in multiple clinical trials targeting several clinical disorders and in the modulation of immune responses. In the present study, we isolated human adipose tissue-derived stem cells (ADSCs) by direct membrane migration method without using enzymatic digestion via collagenase, and tried to extract adequate number of cells for clinical application. Hydroxyapatite-treated nonwoven fabric membrane made up of synthetic macromolecular fiber materials, polyethylene and polyester terephthalene was used. Expansion culture of ADSCs having plastic flask adherent characteristic in serum-free condition was successfully established, and adequate number of cells were obtained for clinical application. They were found to be positive for CD44, CD73, CD90 and CD105 and negative for CD11b, CD34, CD45, CD80 and HLA-DR. The resulting immunological marker profile satisfied the immunophenotype of previously reported MSCs. Also, microscopic findings demonstrated trilineage differentiation into adipogenic, osteogenic and chondrogenic cells as the characteristics of MSCs. The isolation by nonwoven fabric membrane and expanded cells under serum-free condition satisfied the criteria of MSCs, as proposed by the International Society for Cellular Therapy. Our direct membrane migration method without enzyme digestion is useful as ADSCs can be obtained from small pieces of adipose tissue and expanded under serum-free culture condition. This method was considered to be feasible for clinical application.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Zuk PA, Zhu M, Mizuno H, et al. Multilineage cells from human adipose tissue: implications for cell-based therapies. Tissue Eng. 2001;7:211–28.

    Article  CAS  Google Scholar 

  2. Raposio E, Caruana G, Petrella M, et al. A standardized method of isolating adipose-derived stem cells for clinical applications. Ann Plast Surg. 2016;76:124–6.

    Article  CAS  Google Scholar 

  3. da Silva ML, Nardi NB. Methodology, biology and clinical applications of mesenchymal stem cells. Front Biosci Landmark. 2009;14:4281–98.

    Google Scholar 

  4. Wolf D, Wolf AM. Mesenchymal stem cells as cellular immunosuppressants. The Lancet. 2008;371:1553–4.

    Article  Google Scholar 

  5. Doeppner TR, Hermann DK. Mesenchymal stem cells in the treatment of ischemic stroke: progress and possibilities. Stem Cells Cloning. 2010;3:157–63.

    PubMed  PubMed Central  Google Scholar 

  6. Liu Y, Lin L, Zou R, et al. MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis. Cell Cycle. 2018;17:2411–22.

    Article  CAS  Google Scholar 

  7. Tozawa K, Ono-Uruga Y, Yazawa M, et al. Megakaryocytes and platelets from a novel human adiposetissue–derived mesenchymal stem cell line. Blood. 2019;133:633–43.

    Article  CAS  Google Scholar 

  8. Higuchi A, Wang CT, Ling QD, et al. A hybrid-membrane migration method to isolate high-purity adipose-derived stem cells from fat tissues. Sci Rep. 2015;5:10217.

    Article  CAS  Google Scholar 

  9. Lin HR, Heish CW, Liu CH, et al. Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods. Sci Rep. 2017;7:40069.

    Article  CAS  Google Scholar 

  10. https://www.bioind.com/israel/media/wysiwyg/documents/stem-cells/stem-cells-and-cell-therapy.pdf.

  11. Yang Y, Lin H, Shen H, et al. Mesenchymal stem cell-derived extracellular matrix enhances chondrogenic phenotype and cartilage formation by encapsulated chondrocytes in vitro and in vivo. Acta Biomater. 2018;69:71–82.

    Article  CAS  Google Scholar 

  12. Ougushi H, Caplan AI. Stem cell technology and bioceramics: from cell to gene engineering. J Biomed Mater Res. 1999;48(6):913–27.

    Article  Google Scholar 

  13. Brown DV, Filiz G, Daniel PM, et al. Expression of CD133 and CD44 in glioblastoma stem cells correlates with cell proliferation, phenotype stability and intratumor heterogeneity. PLoS ONE. 2017;12:e0172791.

    Article  Google Scholar 

  14. Lee-Sayer SSM, Dougan MN, Cooper J, et al. CD44-mediated hyaluronan binding marks proliferating hematopoietic progenitor cells and promotes bone marrow engraftment. PLoS ONE. 2018;13:e0196011.

    Article  Google Scholar 

  15. Khurana SS, Rieh TE, Moore BD, et al. The hyaluronic acid receptor CD44 coordinates normal and metaplastic gastric epithelial progenitor cell proliferation. J Biol Chem. 2013;288:16085–97.

    Article  CAS  Google Scholar 

  16. Raposio E, Simonacci F, Perrotta RE. Adipose-derived stem cells: comparison between two methods of isolation for clinical applications. Ann Med Surg (Lond). 2017;20:87–91.

    Article  Google Scholar 

  17. Al-Saqi SH, Saliem M, Asikainen S, et al. Defined serum-free media for in vitro expansion of adipose-derived mesenchymal stem cells. Cytotherapy. 2014;16:915–26.

    Article  CAS  Google Scholar 

  18. Tsuji K, Ojima M, Otabe K, Horie M, Koga H, Sekiya I, Muneta T. Effects of different cell-detaching methods on the viability and cell surface antigen expression of synovial mesenchymal stem cells. Cell Transplant. 2017;26:1089–102.

    Article  Google Scholar 

  19. Hendijani F. Explant culture: an advantageous method for isolation of mesenchymal stem cells from human tissues. Cell Prolif. 2017;50:e12334.

    Article  Google Scholar 

  20. Mitchell JB, Mcintosh K, Zvonic S, et al. Immunophenotype of human adipose-derived cells: temporal changes in stromal-associated and stem cell–associated markers. Stem Cells. 2006;24:376–85.

    Article  Google Scholar 

  21. Vyas B, Shah A, Marathe A, et al. Adipose tissue: a natural resource for multipotent mesenchymal stem cells with potential translation to trigerminal layers. Indian J Plast Surg. 2018;51:177–81.

    Article  Google Scholar 

  22. Dominici M, Blanc KL, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. 2016;8:315–7.

    Article  Google Scholar 

Download references

Acknowledgements

We thank Mr. Tatsuo Tanaka, Kyoto WakoPure Chemical Co., Ltd. Kyoto, Japan for his kind procession of hydroxyapatite treatment of non-woven fabric membrane.

Author information

Authors and Affiliations

  1. Division of Cell Therapy Research, U-Medics, Inc, Matsui 8-1, Utano, Uda, Nara, 633-2221, Japan

    Yoshinobu Matsuo & Takahiro Tsujimura

  2. Cellular Biology, Grandsoul Research Institute for Immunology, Inc, Matsui 8-1, Utano, Uda, Nara, 633-2221, Japan

    Yoshinobu Matsuo, Hiromichi Morita & Takahiro Tsujimura

  3. Research and Development Center for Regenerative Medicine, Matsui 8-1, Utano, Uda, Nara, 633-2221, Japan

    Yoshinobu Matsuo, Hiromichi Morita, Hisakazu Yamagishi & Takahiro Tsujimura

  4. Grandsoul Nara Clinic, Matsui 8-1, Utano, Uda, Nara, 633-2221, Japan

    Mitsutoshi Nakamura & Takahiro Tsujimura

  5. Department of Neurosurgery, Nara Medical University, Kashihara, Nara, 634-8522, Japan

    Mitsutoshi Nakamura, Yasuhiro Takeshima & Ichiro Nakagawa

  6. Meiji University of Integrative Medicine, Nantan-City, Kyoto, 629-0392, Japan

    Jiro Imanishi

Authors
  1. Yoshinobu Matsuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hiromichi Morita
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hisakazu Yamagishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Mitsutoshi Nakamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yasuhiro Takeshima
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ichiro Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jiro Imanishi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Takahiro Tsujimura
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yoshinobu Matsuo.

Ethics declarations

Conflict of interest

There is no conflict of interest. There is no grant of funding support for this manuscript.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Matsuo, Y., Morita, H., Yamagishi, H. et al. Isolation of adipose tissue-derived stem cells by direct membrane migration and expansion for clinical application. Human Cell 34, 819–824 (2021). https://doi.org/10.1007/s13577-021-00505-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13577-021-00505-3

Keywords

Search

Navigation