Extraction of human mesenchymal stem cells derived from adipose tissue using various enzymes

Adipose tissue is being used word-wide as a suitable adult stem cell source; the aim of this study was to examine the possibility of using different enzymes to isolate the human mesenchymal stem cells from adipose tissue.

Adipose tissues which were obtained from liposuction surgery were transferred to the laboratory under sterile conditions. All samples obtained from different sources were treated with different enzymes under similar conditions of time and temperature. The cells were assessed for their morphology, density and rate of growth, expression of the specific surface markers, and differentiation abilities.

Regardless of the employed enzymes, cells isolated in the primary cultures showed different morphologies. However following the next subcultures, homogenic populations of cylindrical cells were dominant. In the fourth passage, no differences were observed between different groups in growth rate and the ability of the cells in expression of specific surface markers and differentiation ability into adipose and osteocyte cells. Flow cytometry analysis showed that more than 95% of the cells were positive for the expression of CD73, CD90 and CD105 mesenchymal markers.

Results of this study have demonstrated that wild type collagenase combined with trypsin can be used to isolate the human mesenchymal stem cells from adipose tissue with the same outcome as using pure collagenase, thus reducing the high expense of enzymatic dissociation with pure collagenase.