Abstract 10620: CD271 Expression Enhances Angiogenic Capacity in Human Adipose-Derived Stem Cells

Abstract

Introduction: Therapeutic angiogenesis using stem cell transplantation can be a novel strategy for ischemic disease. Recently, stem cells with gene modifications have been considered as an option to boost therapeutic efficacy. Surface receptor CD271 has emerged as a useful marker for proliferative or angiogenic cell isolation from human organs. We hypothesized CD271 expression on adipose-derived stem cells (ADSCs) would increase their angiogenic capacity. Purpose: We aimed to assess in vitro and in vivo angiogenic capacity of CD271-over expressed human ADSCs. Methods: ADSCs were established from human adipose tissue from healthy subjects and transfected with lentiviral vector for CD271 overexpression (OE-ADSCs). Vector transfection was performed with previously reported method using protamine. Vector without CD271 gene insert was used to prepare control (Cont-ADSCs, Fig.A ). Overexpression of CD271 was evaluated by qPCR and flow cytometry after cell selection using puromycin. For in vivo angiogenic capacity, ADSCs were prelabeled with PKH26 living cell dye, injected via intra-muscular injection into mice subjected to limb ischemia model, and evaluated by immunohistochemistry of ischemic limb (gastrocnemius). Results: We found that mRNA expression of CD271 was 10,000-fold higher in OE-ADSCs than in Cont-ADSCs. Consistently, over 90% of OE-ADSCs expressed CD271 protein. ( Fig.B ). Overexpression of CD271 on ADSCs increased mRNA expression of angiogenic factors such as VEGFA and CXCL12 . Furthermore, mRNA expression of TIMP2, anti-angiogenic factor, was significantly decreased in OE-ADSCs ( Fig.C ). In cell therapy, we demonstrated that administration of OE-ADSCs enhanced in vivo angiogenic capacity evaluated by capillary density. Notably, ADSC engraftment capacity was also augmented in OE-ADSCs ( Fig.D ). Conclusion: These findings highlight the important role of CD271 expression on ADSCs for their angiogenic gene profile and cell therapy effects.