Serial Cultivation of an MSC-Like Cell Line with Enzyme-Free Passaging Using a Microporous Titanium Scaffold

Autor: Yukihiko Sakisaka, Hiroshi Ishihata, Kentaro Maruyama, Eiji Nemoto, Shigeki Chiba, Masaru Nagamine, Hiroshi Hasegawa, Takeshi Hatsuzawa, Satoru Yamada
Jazyk: angličtina
Rok vydání: 2023
Předmět:
Zdroj: Materials, Vol 16, Iss 3, p 1165 (2023)
Druh dokumentu: article
ISSN: 1996-1944
DOI: 10.3390/ma16031165
Popis: In vitro studies on adherent cells require a process of passage to dissociate the cells from the culture substrate using enzymes or other chemical agents to maintain cellular activity. However, these proteolytic enzymes have a negative influence on the viability and phenotype of cells. The mesenchymal stem cell (MSC)-like cell line, C3H10T1/2, adhered, migrated, and proliferated to the same extent on newly designed microporous titanium (Ti) membrane and conventional culture dish, and spontaneous transfer to another substrate without enzymatic or chemical dissociation was achieved. The present study pierced a 10 μm-thick pure Ti sheet with 25 μm square holes at 75 μm intervals to create a dense porous structure with biomimetic topography. The pathway of machined holes allowed the cells to access both sides of the membrane frequently. In a culture with Ti membranes stacked above- and below-seeded cells, cell migration between the neighboring membranes was confirmed using the through-holes of the membrane and contact between the membranes as migration routes. Furthermore, the cells on each membrane migrated onto the conventional culture vessel. Therefore, a cell culture system with enzyme-free passaging was developed.
Databáze: Directory of Open Access Journals
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