Tissue Eng Regen Med.  2019 Feb;16(1):19-28. 10.1007/s13770-018-00174-8.

Tissue Regeneration of Human Mesenchymal Stem Cells on Porous Gelatin Micro-Carriers by Long-Term Dynamic In Vitro Culture

Affiliations
  • 1Department of Chemical and Biomolecular Engineering, Seoul National University of Science and Technology, 232 Gongneung-ro, Nowon-gu, Seoul 11811, Republic of Korea. insup@seoultech.ac.kr
  • 2Convergence Institute of Biomedical Engineering and Biomaterials, Seoul National University of Science and Technology, 232 Gongneung-ro, Nowon-gu, Seoul 11811, Republic of Korea.

Abstract

BACKGROUND
Tissue engineering is a multidisciplinary field which attracted much attention in recent years. One of the most important issue in tissue engineering is how to obtain high cell numbers and tissue regeneration while maintaining appropriate cellular characteristics in vitro for restoring damaged or dysfunctional body tissues and organs. These demands can be achieved by the use of three dimensional (3D) dynamic cultures of cells combined with cell-adhesive micro-carriers.
METHODS
In this study, human mesenchymal stem cells (hMSCs) were cultured in a wave-bioreactor system for up to 100 days, after seeding on Cultisphere-S porous gelatin micro-carriers. Cell counting was performed at the time points of 7, 12, 17, 31 days and compared to those of hMSCs cultured under static condition. Higher growth and proliferation rates was achieved in wave-type dynamic culture, when cell culture continued to day 31. A scanning electron microscope (SEM) photographs, both live and dead and MTT assays were taken to confirm the survival and distribution of cells on porous gelatin micro-carrier surfaces. The results of histological stains such as hematoxylin and eosin, Masson's trichrome, Alcian blue and Alizarin red S also showed improved proliferation and tissue regeneration of hMSCs on porous gelatin micro-carriers.
CONCLUSION
The experimental results demonstrated the effect and importance of both micro-carriers and bioreactor in hMSC expansion on cell proliferation and migration as well as extracellular matrix formation on the superficial and pore surfaces of the porous gelatin micro-carriers, and then their inter-connections, leading to tissue regeneration.

Keyword

Dynamic culture; Waving type bioreactor; Mesenchymal stem cells; Gelatin micro-carrier

MeSH Terms

Alcian Blue
Bioreactors
Cell Count
Cell Culture Techniques
Cell Proliferation
Coloring Agents
Eosine Yellowish-(YS)
Extracellular Matrix
Gelatin*
Hematoxylin
Humans*
In Vitro Techniques*
Mesenchymal Stromal Cells*
Regeneration*
Tissue Engineering
Alcian Blue
Coloring Agents
Eosine Yellowish-(YS)
Gelatin
Hematoxylin
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