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Stem Cells International
Volume 2016 (2016), Article ID 3274054, 9 pages
http://dx.doi.org/10.1155/2016/3274054
Research Article

Evaluation of Tissue Homogenization to Support the Generation of GMP-Compliant Mesenchymal Stromal Cells from the Umbilical Cord

1St. Louis Cord Blood Bank/Cellular Therapy Laboratory, SSM Health Cardinal Glennon Children’s Hospital, St. Louis, MO 63110, USA
2Department of Pediatrics, Saint Louis University School of Medicine, St. Louis, MO 63104, USA
3SSM Health St. Mary’s Hospital, St. Louis, MO 63117, USA
4Department of Obstetrics, Gynecology and Women’s Health, Saint Louis University School of Medicine, St. Louis, MO 63104, USA

Received 23 September 2015; Revised 2 December 2015; Accepted 14 December 2015

Academic Editor: Peter Czermak

Copyright © 2016 Ryan J. Emnett et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Recent studies have demonstrated that the umbilical cord (UC) is an excellent source of mesenchymal stromal cells (MSCs). However, current protocols for extracting and culturing UC-MSCs do not meet current good manufacturing practice (cGMP) standards, in part due to the use of xenogeneic reagents. To support the development of a cGMP-compliant method, we have examined an enzyme-free isolation method utilizing tissue homogenization (t-H) followed by culture in human platelet lysate (PL) supplemented media. The yield and viability of cells after t-H were comparable to those obtained after collagenase digestion (Col-D). Importantly, kinetic analysis of cultured cells showed logarithmic growth over 10 tested passages, although the rate of cell division was lower for t-H as compared to Col-D. This slower growth of t-H-derived cells was also reflected in their longer population doubling time. Interestingly, there was no difference in the expression of mesenchymal markers and trilineage differentiation potential of cells generated using either method. Finally, t-H-derived cells had greater clonogenic potential compared to Col-D/FBS but not Col-D/PL and were able to maintain CFU-F capacity through P7. This bench scale study demonstrates the possibility of generating therapeutic doses of good quality UC-MSCs within a reasonable length of time using t-H and PL.