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

Chen, Shyi-Gen MD; Tseng, Pei-Chi PhD; Huang, Chi-Hsuan BA; Shen, Pei-Chen MS; Lo, Wei-Yu PhD; Chan, James Yi-Hsin MD, PhD

doi: 10.1097/SAP.0000000000000704
Research Papers
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Background 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.

Methods 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.

Results 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.

Conclusions 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.

From the *Plastic and Reconstructive Surgery, Tri-Service General Hospital, National Defense Medical Center; †MSC Laboratory, R&D Department, HealthBanks Biotech Co, Ltd; and ‡Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan.

Received October 19, 2015, and accepted for publication, after revision October 26, 2015.

S.-G.C. and P.-C.T. are co–first authors; these authors contributed equally to this work.

Conflicts of interest and sources of funding: Financial support for this work was provided by MSC Laboratory; R&D Department; HealthBanks Biotech Co, Ltd, Taiwan; National Defense Medical Center and Tri-Service General Hospital, ROC (TSGH-C102-006-008-013-S01, TSGH-C102-006-008-013-S04); and the Ministry of Science and Technology (102-2314-B016-010).

Reprints: Wei-Yu Lo, PhD, R&D Department, HealthBanks Biotech Co, Ltd. 5F, No 25, Sec. 4, Ren Ai Road, Taipei 106, Taiwan. E-mail: wylo@healthbanks.com.tw; James Yi-Hsin Chan, MD, PhD, Department of Microbiology and Immunology; Department of Family and Community Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei 114, Taiwan, No 161, Sec. 6, Minquan E. Road, Taipei 114, Taiwan. E-mail: jchan9473@gmail.com.

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