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Protocol of mesenchymal stem cell inoculation to nonwoven fabric scaffold

Authors
  • Fu, Bo1
  • Yamada, Kyosuke1
  • Fujiwara, Masashi1
  • Iwai, Ryosuke2, 3
  • Takagi, Mutsumi1
  • 1 Hokkaido TechUniversity, Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Kita-ku, N13W8, Sapporo, 060-8628, Japan , Sapporo (Japan)
  • 2 Okayama University of Science, Research Institute of Technology, 1-1 Ridaicho, Kita-ku, Okayama-shi, 700-0005, Japan , Okayama-shi (Japan)
  • 3 National Cerebral and Cardiovascular Center, Research Institute, 5-7-1 Fujishiro-dai, Suita, Osaka, 565-8565, Japan , Suita (Japan)
Type
Published Article
Journal
Cytotechnology
Publisher
Springer-Verlag
Publication Date
May 23, 2019
Volume
71
Issue
3
Pages
743–750
Identifiers
DOI: 10.1007/s10616-019-00320-7
Source
Springer Nature
Keywords
License
Yellow

Abstract

To obtain a large number of human mesenchymal stem cell (hMSCs) for allograft, nonwoven fabrics (NWF) were used as a cell culture scaffold. NWF are three-dimensional fiber aggregates formed by heat bonding and have a high surface area for cell adhesion and elongation. Inoculation hMSC was done to a center of NWF disc (diameter, 15.1 mm; depth, 0.1 mm). A cell suspension inoculum had a volume of 10 µL, which was close to the void volume of the disc, and resulted in a high initial (24 h) cell adhesion efficiency. Use of green fluorescent protein expressing rat MSCs and fluorescence microscopy revealed that adding an additional 10 μL of medium at 0–2 h after the cell inoculation made the initial horizontal distribution of cells in the NWF disc more uniform. Addition of 10 μL of the medium after 1 and 2 h of hMSC inoculation (0.15 × 103 cells/cm2 NWF-fiber) markedly increased the final cell density (21 days) from 2.48 to 7.45 × 103 cells/cm2 NWF-fiber and fold increase in cell density by 16–48-fold. In conclusion, the addition of an additional medium after inoculation made the initial cells distribution in NWF more uniform, which might result in higher final cell density.

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