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Differentiation of human mesenchymal stem cell spheroids under microgravity conditions

Wolfgang H Cerwinka15*, Starlette M Sharp2, Barbara D Boyan3, Haiyen E Zhau4, Leland W K Chung4 and Clayton Yates26*

Author Affiliations

1 Children’s Healthcare of Atlanta, Emory University School of Medicine, 5445Meridian Mark Road, Suite 420, Atlanta, GA 30342, USA

2 Department of Biology and Center for Cancer Research, Tuskegee University, Carver Research Building, Tuskegee Institute, AL 36088, USA

3 The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 313 Ferst Drive Atlanta, Atlanta, GA 30332, USA

4 Samuel-Oschin Comprehensive Cancer Institute, Cedars -Sinai Medical Center, 8750 Beverly Blvd., Atrium 103, Los Angeles, CA 90048, USA

5 Georgia Pediatric Urology, 5445 Meridian Mark Rd, Suite 420, Atlanta, GA 30342, USA

6 Tuskegee University, Carver Research Bld. Rm 22, Tuskegee, AL 36088, USA

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Cell Regeneration 2012, 1:2  doi:10.1186/2045-9769-1-2

Published: 28 June 2012


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.

Mesenchymal stem cell; RWV culture; Tissue engineering; Differentiation; Osteogenesis