Protocol of mesenchymal stem cell inoculation to nonwoven fabric scaffold
| Autor: | Bo Fu, Masashi Fujiwara, Mutsumi Takagi, Kyosuke Yamada, Ryosuke Iwai |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Nonwoven fabric Chemistry Clinical Biochemistry Cell Mesenchymal stem cell Biomedical Engineering Bioengineering Cell Biology JAACT Issue Green fluorescent protein 03 medical and health sciences 030104 developmental biology 0302 clinical medicine medicine.anatomical_structure Cell culture 030220 oncology & carcinogenesis medicine Biophysics Fluorescence microscope Elongation Cell adhesion Biotechnology |
| Zdroj: | Cytotechnology |
| Popis: | 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 × 10(3) cells/cm(2) NWF-fiber) markedly increased the final cell density (21 days) from 2.48 to 7.45 × 10(3) cells/cm(2) 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. |
| Databáze: | OpenAIRE |
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