Skip to main content
SpringerLink
Log in

Effects of agitation rate on aggregation during beads-to-beads subcultivation of microcarrier culture of human mesenchymal stem cells

  • Original Article
  • Published:
Cytotechnology Aims and scope Submit manuscript

Abstract

With the aim to utilize human mesenchymal stem cells (hMSCs) grown in large scale for regenerative medicine, effects of agitation rate on aggregation during beads-to-beads subcultivation of microcarrier culture of hMSCs were studied. hMSCs could attach and grew on surface-type microcarriers of Cytodex 1, whereas almost no cell elongation and growth were observed on porous type microcarriers of Cytopores. The percentages of aggregated Cytodex 1 microcarriers at an agitation rate of 60 and 90 rpm were lower than that at 30 rpm, which was the lowest agitation rate necessary for the suspension of Cytodex 1 microcarriers, and the cells grew fastest at 60 rpm. hMSC could be subcultivated on Cytodex 1 by the beads-to-beads method at both 30 and 60 rpm without trypsinization. However, agitation at 60 rpm resulted in a markedly lower percentage of aggregated microcarriers not only before but also after subcultivation. The percentages of CD90- and CD166-positive cells among cells grown on Cytodex 1 at 60 rpm (91.5 and 87.6 %) were comparable to those of cells grown in the pre-culture on dishes. In conclusion, hMSCs could be subcultivated on Cytodex 1 by beads-to-beads method maintaining the expressions of the cell surface antigens CD90 and CD166, while adjusting agitation rate could decrease the microcarrier aggregation.

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.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  • Borys MC, Papoutsakis ET (1992) Formation of bridges and large cellular clumps in CHO-cell microcarrier cultures: effects of agitation, dimethyl sulfoxide and calf serum. Cytotechnology 8:237–248

    Article  CAS  Google Scholar 

  • Chua S, Yuen CM, Leu S, Lin YC, Sun CK, Yip HK (2010) Adipose-derived mesenchymal stem cells markedly attenuate brain infarct size and improve neurological function in rats. Eur Heart J 31:80

    Google Scholar 

  • Ferrari C, Balandras F, Guedon E, Olmos E, Chevalot I (2012) Limiting cell aggregation during mesenchymal stem cell expansion on microcarriers. Biotechnol Prog 28:780–787

    Article  CAS  Google Scholar 

  • Frauenschuh S, Reichmann E, Ibold Y, Goetz PM, Sittinger M, Ringe J (2007) A microcarrier-based cultivation system for expansion of primary mesenchymal stem cells. Biotechnol Prog 23:187–193

    Article  CAS  Google Scholar 

  • Malda J, Van Blitterswijk CA, Grojec M, Martens DE, Tramper J, Riesle J (2003) Expansion of bovine chondrocytes on microcarriers enhances redifferentiation. Tissue Eng 9:939–948

    Article  CAS  Google Scholar 

  • Nienow AW (2006) Reactor engineering in large scale animal cell culture. Cytotechnology 50:9–33

    Article  CAS  Google Scholar 

  • Sanford KK, Earle WR, Evans VJ, Waltz HK, Shannon JE (1950) The measurement of proliferation in tissue cultures by enumeration of cell nuclei. Anat Rec 106:243

    Google Scholar 

  • Sato Y, Wakitani S, Takagi M (2013) Xeno-free and shrinkage-free preparation of scaffold-free cartilage-like disc-shaped cell sheet using human bone marrow mesenchymal stem cells. J Biosci Bioeng 116:734–739

    Article  CAS  Google Scholar 

  • Schop D, Janssen FW, de Bruijn JD, Van Dijkhuizen-Radersma R (2008) Expansion of mesenchymal stem cells using a microcarrier-based cultivation system: growth and metabolism. J Tissue Eng Regen Med 2:126–135

    Article  CAS  Google Scholar 

  • Shiragami N, Honda H, Unno H (1993) Anchorage-dependent animal cell culture by using a porous microcarrier. Bioprocess Eng 8:295–299

    Article  CAS  Google Scholar 

  • Takagi M, Okumura H, Okada T, Kobayashi N, Kiyota T, Ueda K (1994) An oxygen supply strategy for the large-scale production of tissue plasminogen activator by microcarrier cell culture. J Ferment Bioeng 77:301–306

    Article  CAS  Google Scholar 

  • Takagi M, Sasaki T, Yoshida T (1999) Spatial development of the cultivation of a bone marrow stromal cell line in porous carriers. Cytotechnology 31:225–231

    Article  CAS  Google Scholar 

  • Wang Y, Ouyang F (1999) Bead-to-bead transfer of Vero cells in microcarrier culture. Cytotechnology 31:221–224

    Article  CAS  Google Scholar 

  • Yamahara K, Taguchi A, Soma T, Ogawa H, Ikeda T, Yoshimatsu J (2014) Clinical application of human amnion-derived mesenchymal stem cells for the treatment of acute GVHD. Placenta 35:A6

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido University, Kita-ku, N13W8, Sapporo, 060-8628, Japan

    Iori Takahashi, Keigo Sato & Mutsumi Takagi

  2. School of Health and Sports Sciences, Mukogawa Women’s University, 6-46 Ikebiraki, Nishinomiya, Hyogo, 663-8558, Japan

    Hisashi Mera

  3. Foundation for Biomedical Research and Innovation, International Medical Device Alliance, 1-6-5 Minatojima Minamimachi, Chuo-Ku Kobe, Hyogo, 650-0047, Japan

    Hisashi Mera

  4. Graduate School of Biomedical Sciences, Hiroshima University, Hiroshima, 1-2-3 Kasumi, Minami-ku, Hiroshima, 739-8553, Japan

    Shigeyuki Wakitani

Authors
  1. Iori Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Keigo Sato
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Hisashi Mera
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shigeyuki Wakitani
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mutsumi Takagi
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mutsumi Takagi.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Takahashi, I., Sato, K., Mera, H. et al. Effects of agitation rate on aggregation during beads-to-beads subcultivation of microcarrier culture of human mesenchymal stem cells. Cytotechnology 69, 503–509 (2017). https://doi.org/10.1007/s10616-016-9999-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10616-016-9999-5

Keywords

Search

Navigation