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Fabrication of size-controllable human mesenchymal stromal cell spheroids from micro-scaled cell sheets.

08:00 EDT 16th May 2019 | BioPortfolio

Summary of "Fabrication of size-controllable human mesenchymal stromal cell spheroids from micro-scaled cell sheets."

Recently, stem cell spheroids have been actively studied for use in tissue regeneration. In this study, we report a method for the fabrication of size-controllable stem cell spheroids in different sizes from micro-scaled cell sheets (μCS) using thermosensitive hydrogels and investigated their effects on stem cell function. Mesenchymal stem cells isolated from different tissues such as human turbinate tissue, bone marrow, and adipose tissue were adhered selectively to each micro-pattern (squares with widths of 100 and 400 μm) on the surface of the hydrogel and formed μCS. The diameters of the spheroids were modulated by the size of the patterns (45±5 and 129±4 μm in diameter for the 100 and 400 μm micro-patterns, respectively) and the seeding density (129±4, 149±6, and 163±6 μm for 5.0, 10.0, and 15.0 × 10<sup>4</sup> cells/cm<sup>2</sup>, respectively, on 400-μm micro-pattern). In addition, the spheroids were successfully fabricated regardless of stem cell origin, and the diameter of the spheroids was also affected by cell spreading area on a cell culture dish. Stemness markers were highly expressed in the spheroids regardless of the spheroid size. Furthermore, an increase in E-cadherin and decrease in N-cadherin gene expression showed the stable formation of spheroids of different sizes. Gene expression levels of hypoxia inducible factors and secretion of vascular endothelial growth factor (VEGF) were increased (13.2±1.4, 325±83.4 and 534.3±121.5 pg/ng DNA in a monolayer, and 100- and 400-μm micro-patterned spheroids, respectively) proportional to the diameters of the spheroids. The size of spheroids were maintained even after injection, cryopreservation and 7 days of suspension culture with high viability (~90%). In conclusion, this novel technique to fabricate spheroids with controlled size could be widely applied in various applications that require a controlled size in regenerative medicine.

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Name: Biofabrication
ISSN: 1758-5090
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