Skip to main content
SpringerLink
Log in

Endothelial cell behavior inside myoblast sheets with different thickness

  • Original Research Paper
  • Published:
Biotechnology Letters Aims and scope Submit manuscript

Abstract

Using a cell sheet stacking method, we developed an in vitro culture system in which green fluorescent protein expressing human umbilical vein endothelial cells (GFP-HUVECs) were cultured under human skeletal muscle myoblast (HSMM) sheets with different layer numbers. Our aim in developing this system was to examine the different endothelial behaviors in the cell sheet. During 96 h of incubation, in monolayer HSMM sheet, HUVECs quickly reached the top of the cell sheet and detached. In three-layered HSMM sheet, HUVECs also migrated to the top layer and formed island-shaped aggregates. In five-layered HSMM sheet, HUVECs migrated into the middle of the cell sheet and formed net-shaped aggregates. In seven-layered HSMM sheet, HUVECs migrated in the basal of the cell sheet and formed sparse net-shaped aggregates. The thickness of the HSMM sheet, which can be controlled by the layer number of the cell sheet, is therefore an important parameter that affects the migration time, encounters, localization, and morphology of HUVECs inside the HSMM sheet.

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

Similar content being viewed by others

References

  • Al Attar N, Razak AB, Scorsin M (2002) Cellular transplantation: new horizons in the surgical management of heart failure. J Roy Coll Surg Edinb 47:749–752

    PubMed  CAS  Google Scholar 

  • Almagro S, Durmort C, Chervin-Petinot A, Heyraud S, Dubois M, Lambert O, Maillefaud C, Hewat E, Schaal JP, Huber P et al (2010) The motor protein myosin-X transports VE-cadherin along filopodia to allow the formation of early endothelial cell–cell contacts. Mol Cell Biol 30:1703–1717

    Article  PubMed  CAS  Google Scholar 

  • Asakawa N, Shimizu T, Tsuda Y, Sekiya S, Sasagawa T, Yamato M, Fukai F, Okano T (2010) Pre-vascularization of in vitro three-dimensional tissues created by cell sheet engineering. Biomaterials 31:3903–3909

    Article  PubMed  CAS  Google Scholar 

  • Chowdhury SR, Muneyuki Y, Takezawa Y, Kino-oka M, Saito A, Sawa Y, Taya M (2010) Growth and differentiation potentials in confluent state of culture of human skeletal muscle myoblasts. J Biosci Bioeng 109:310–313

    Article  PubMed  CAS  Google Scholar 

  • Hassink RJ, de la Riviere AB, Mummery CL, Doevendans PA (2003) Transplantation of cells for cardiac repair. J Am Coll Cardiol 41:711–717

    Article  PubMed  Google Scholar 

  • Kino-oka M, Ngo TX, Nagamori E, Takezawa Y, Miyake Y, Sawa Y, Saito A, Shimizu T, Okano T, Taya M (2012) Evaluation of vertical cell fluidity in a multilayered sheet of skeletal myoblasts. J Biosci Bioeng 113:128–131

    Article  PubMed  CAS  Google Scholar 

  • Martins GG, Kolega J (2006) Endothelial cell protrusion and migration in three-dimensional collagen matrices. Cell Motil Cytoskelet 63:101–115

    Article  Google Scholar 

  • Memon IA, Sawa Y, Fukushima N, Matsumiya G, Miyagawa S, Taketani S, Sakakida SK, Kondoh H, Aleshin AN, Shimizu T et al (2005) Repair of impaired myocardium by means of implantation of engineered autologous myoblast sheets. J Thorac Cardiovasc Surg 130:1333–1341

    Article  PubMed  Google Scholar 

  • Miyagawa S, Roth M, Saito A, Sawa Y, Kostin S (2011) Tissue-engineered cardiac constructs for cardiac repair. Ann Thorac Surg 91:320–329

    Article  PubMed  Google Scholar 

  • Nagamori E, Ngo TX, Tekezawa Y, Saito A, Sawa Y, Shimizu T, Okano T, Taya M, Kino-oka M (2013) Network formation through active migration of human vascular endothelial cells in a multilayered skeletal myoblast sheet. Biomaterials 34:662–668

    Article  PubMed  CAS  Google Scholar 

  • Perez-Ilzarbe M, Agbulut O, Pelacho B, Ciorba C, Jose-Eneriz ES, Desnos M, Hagege AA, Aranda P, Andreu EJ, Menasche P et al (2008) Characterization of the paracrine effects of human skeletal myoblasts transplanted in infarcted myocardium. Eur J Heart Fail 10:1065–1072

    Article  PubMed  CAS  Google Scholar 

  • Sawa Y, Miyagawa S, Sakaguchi T, Fujita T, Matsuyama A, Saito A, Shimizu T, Okano T (2012) Tissue engineered myoblast sheets improved cardiac function sufficiently to discontinue LVAS in a patient with DCM: report of a case. Surg Today 42:181–184

    Article  PubMed  Google Scholar 

  • Sekiya N, Matsumiya G, Miyagawa S, Saito A, Shimizu T, Okano T, Kawaguchi N, Matsuura N, Sawa Y (2009) Layered implantation of myoblast sheets attenuates adverse cardiac remodeling of the infarcted heart. J Thorac Cardiovasc Surg 138:985–993

    Article  PubMed  Google Scholar 

  • Sekiya S, Muraoka M, Sasagawa T, Shimizu T, Yamato M, Okano T (2010) Three-dimensional cell-dense constructs containing endothelial cell-networks are an effective tool for in vivo and in vitro vascular biology research. Microvasc Res 80:549–551

    Article  PubMed  Google Scholar 

  • Wood W, Martin P (2002) Structures in focus—filopodia. Int J Biochem Cell Biol 34:726–730

    Article  PubMed  CAS  Google Scholar 

  • Zammaretti P, Jaconi M (2004) Cardiac tissue engineering: regeneration of the wounded heart. Curr Opin Biotech 15:430–434

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This study was supported by the Japan Society for the Promotion of Science (JSPS) through the “Funding Program for World-Leading Innovative R&D on Science and Technology (FIRST Program)” initiated by the Council for Science and Technology Policy (CSTP) and the Japan Science and Technology Agency (JST).

Author information

Authors and Affiliations

  1. Department of Biotechnology, Graduate School of Engineering, Osaka University, 2-1 Yamada-oka, Suita, Osaka, 565-0871, Japan

    Eiji Nagamori & Masahiro Kino-oka

  2. Division of Chemical Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka, 560-8531, Japan

    Trung Xuan Ngo & Masahito Taya

  3. Institute of Advanced Biomedical Engineering and Science, Tokyo Women’s Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo, 162-8666, Japan

    Tetsutaro Kikuchi, Tatsuya Shimizu & Teruo Okano

Authors
  1. Trung Xuan Ngo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Eiji Nagamori
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tetsutaro Kikuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Tatsuya Shimizu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Teruo Okano
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Masahito Taya
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Masahiro Kino-oka
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Masahiro Kino-oka.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (MPG 4318 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ngo, T.X., Nagamori, E., Kikuchi, T. et al. Endothelial cell behavior inside myoblast sheets with different thickness. Biotechnol Lett 35, 1001–1008 (2013). https://doi.org/10.1007/s10529-013-1174-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10529-013-1174-x

Keywords

Search

Navigation