Selective Criteria and Markers in Adipose-Derived Stromal Cells Collection Quality and Expansion Potency.

Abstract

Adipose tissue-derived stromal cells (ADSCs) have been extensively used in clinical trials for various therapeutic applications. However, there is a paucity of selective criteria regarding collection and expansion procedures. The purpose of this study was to investigate the effect of liposuction and donor age on ADSCs and to assess the criteria and markers for ADSC long-term expansion potential. Adipose tissues were collected using syringe liposuction, water-jet, or ultrasonic techniques. Tissue/cell viability was evaluated using the XTT assay. CD34 and SSEA-4 expression were examined using flow cytometry. SOX2 gene expression was estimated using quantitative polymerase chain reaction, and Nile-red staining was performed to evaluate the adipogenesis potency during ADSC expansion. The lipoaspirates obtained from syringe and ultrasonic liposuction methods were superior to those of the water-jet method in stromal vascular fraction yield and durability during temporary storage. SSEA-4, SOX2 expression, and adipogenesis potency of early-passage ADSCs were significantly correlated with the P15 cumulative population doublings data. CD34 expression was strongly correlated with P0 ADSC yield and doubling time. Tissue viability, P0 ADSC CD34⁺ percentage, and P15 cumulative population doublings were decreased along with donor age. This study established criteria and markers to determine whether lipoaspirate tissue and cultured ADSCs are suitable for further large-scale expansion and allogenic universal cell banking.