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Differentiation of human mesenchymal stem cell spheroids under microgravity conditions

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Author(s): Cerwinka Wolfgang H | Sharp Starlette M | Boyan Barbara D | Zhau Haiyen E | Chung Leland W K | Yates Clayton

Journal: Cell Regeneration
ISSN 2045-9769

Volume: 1;
Issue: 1;
Start page: 2;
Date: 2012;
Original page

Keywords: Mesenchymal stem cell | RWV culture | Tissue engineering | Differentiation | Osteogenesis

ABSTRACT
Abstract To develop and characterize a novel cell culture method for the generation of undifferentiated and differentiated human mesenchymal stem cell 3D structures, we utilized the RWV system with a gelatin-based scaffold. 3 × 106 cells generated homogeneous spheroids and maximum spheroid loading was accomplished after 3 days of culture. Spheroids cultured in undifferentiated spheroids of 3 and 10 days retained expression of CD44, without expression of differentiation markers. Spheroids cultured in adipogenic and osteogenic differentiation media exhibited oil red O staining and von Kossa staining, respectively. Further characterization of osteogenic lineage, showed that 10 day spheroids exhibited stronger calcification than any other experimental group corresponding with significant expression of vitamin D receptor, alkaline phosphatase, and ERp60 . In conclusion this study describes a novel RWV culture method that allowed efficacious engineering of undifferentiated human mesenchymal stem cell spheroids and rapid osteogenic differentiation. The use of gelatin scaffolds holds promise to design implantable stem cell tissue of various sizes and shapes for future regenerative treatment.
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RPA Switzerland

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