Comparative characterization of frozen-thawed CD146+ and CD146- subsets of CD73+CD90+CD105+CD34+ human ASCs.

Abstract

Mesenchymal stem cells are currently used to treat several diseases. Populations of putative stem cells found in the adipose tissue (ASCs) have been shown to possess particularly enhanced functionalities. Nonetheless, there is lack of evidence that evaluates the effects of cryopreservation techniques on well-defined functional ASC populations characterized by immunophenotypical repertoire. We therefore embarked a study to compare the frozen-thawed ASC subsets: CD73+CD90+CD105+CD34+CD146-(CD34+CD146), CD73+CD90+CD105+CD34+CD146+(CD34+CD146+), and CD73+CD90+CD105+CD34+(CD34+). We assessed their characterization in different functional assays. The ASC immunophenotypical subsets-purified by a flow cytometry sorting technique-were frozen in liquid nitrogen. After a period, they were thawed to examine their differentiation ability, colony-forming units, viability, and growth rate. We confirmed that inside the primary cell culture system, the proportion of CD34+, CD34+CD146-, and CD34+CD146+ took up 80%, 62%, and 19% on average, respectively. All populations could be frozen and stored in liquid nitrogen with retention of more than 85% of cell viability and displayed comparable stemness characteristics. Most importantly, the CD34+CD146+ subpopulation displayed a higher proliferation rate than other groups. Our data demonstrated that the frozen-thawed CD34+CD146+ cells might represent a promising source for autologous cellular-based therapy. These findings set the basis for ASC subpopulations-based application in future potential clinical settings.