5.08 A Novel Method of Human Adipose-derived Stem Cell Isolation With Resultant Increased Cell Yield

Posted on December 22, 2014

R. Tevlin1,2, A. Mc Ardle1,2, E. Seo2, M. T. Chung1, K. Paik1, E. R. Zielins1, D. Atashroo1, M. S. Hu1, G. G. Walmsley1,2, S. Li1, C. K. Chan2, D. C. Wan1, M. T. Longaker1,2  1Stanford University,Plastic Surgery,Palo Alto, CA, USA 2Stanford University,Institute for Stem Cell Biology and Regenerative Medicine,Palo Alto, CA, USA

Introduction:

Adipose tissue represents an abundant and easily accessible source of multipotent cells which may serve as excellent building blocks for tissue engineering. We have developed a new protocol for isolating adipose-derived stromal cells (ASC) from human lipoaspirate. The aim of this study was to compare our new method for ASC isolation to standard protocol (Zuk et al, 2002).

Methods:

Human ASC isolation was performed using two methods to compare cell yield, cell viability and cell proliferation. The osteogenic and adipogenic potential of ASCs isolated using both protocols was assessed in vitro and gene expression analysis was performed. To assess the ability of isolated ASCs to generate bone in vivo, we created critical sized calvarial defects in mice which we treated with ASCs loaded onto HA-PLGA scaffolds.

Results:
We have developed a novel protocol which results in a 10-fold increased yield of ASCs compared to conventional technique. Resulting ASCs harvested using our new protocol demonstrate significantly increased cell viability and proliferation in vitro (p<0.05). Additionally, ASCs isolated using our new protocol demonstrated significantly enhanced osteogenic and adipogenic differentiation capacity in vitro (p<0.05). ASCs harvested using our new protocol have an equal ability to heal critical sized calvarial defects in mice.

Conclusion:

We have developed a protocol that maximizes the yield of ASCs derived from lipoaspirate. We demonstrate that ASCs harvested using this method have an increased osteogenic and adipogenic potential in vitro and are equivalent to ASCs isolated using the conventional protocol in terms of their ability to generate bone in vivo.

 

Reference: 

Zuk PA, Zhu M, Ashjian P, De Ugarte DA, Huang JI, Mizuno H et al. (2002). Human adipose tissue is a source of multipotent stem cells. Molecular biology of the cell 13(12):4279-4295.