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Author Notes:

Correspondence: wcerwin@emory.edu Children’s Healthcare of Atlanta, Emory University School of Medicine, 544 5 Meridian Mark Road, Suite 420, Atlanta, GA 30342, USA; cyates@mytu.tuskegee.edu Georgia Pediatric Urology, 5445 Meridian Mark Rd, Suite 420, Atlanta, GA 30342, USA

WC and CY conceived, designed, and performed the experiments and analyzed the data.

SS, BB HZ performed experiments and data analysis. LWKC, WC, and CY wrote the manuscript.

All authors have read and approve the final manuscript.

We thank Dr. Carlos Abramowsky and Carla Shoffeit from the Department of Pediatric Pathology at Children’s Healthcare of Atlanta for their help in tissue preparation and staining.

The authors declare that they have no competing interests.


Research Funding:

This grant was supported by PO-1 CA-098912, NIH/RCMI G12 RR03059-21A1NIH/NCHD, NIH/NCI 1 U54 CA118623.


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

Differentiation of human mesenchymal stem cell spheroids under microgravity conditions


Journal Title:

Cell Regeneration


Volume 1, Number 1


, Pages 2-2

Type of Work:

Article | Final Publisher PDF


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 × 10 6 cells generated homogeneous spheroids and maximum spheroid loading was accomplished after 3days 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, sh owed that 10 day spheroids exhibited stronger calcification than any other experimental group corresponding with significant expression of vitamin D receptor, alkaline phosphatase, and ERp 60. 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.

Copyright information:

© 2012 Cerwinka et al.

This is an Open Access work distributed under the terms of the Creative Commons Attribution 2.0 Generic License (http://creativecommons.org/licenses/by/2.0/).

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