A novel three-dimensional adipose-derived stem cell cluster for vascular regeneration in ischemic tissue

Abstract

Background aims. Stem cells are one of the most powerful tools in regeneration medicine. However, many limitations remain regarding the use of adult stem cells in clinical applications, including poor cell survival and low treatment efficiency. We describe an innovative three-dimensional cell mass (3DCM) culture that is based on cell adhesion (basic fibroblast growth factor immobilized substrate) and assess the therapeutic potential of 3DCMs composed of human adipose tissue derived stromal cells (hASCs). Methods. For formation of a 3DCM, hASCs were cultured on a substrate with immobilized fibroblast growth factor-2. The angiogenic potential of 3DCMs was determined by immunostaining, fluorescence-activated cell sorting and protein analysis. To evaluate the vasculature ability and improved treatment efficacy of 3DCMs, the 3DCMs were intramuscularly injected into the ischemic limbs of mice. Results. The 3DCMs released various angiogenic factors (eg, vascular endothelial growth factor and interleukin-8) and differentiated into vascular cells within 3 days in normal medium. Blood vessel and tissue regeneration was clearly observed through visual inspection in the 3DCM-injected group. hASC injection slowed limb necrosis after treatment, but 50% of the mice ultimately had limb loss within 28 days. Most mice receiving 3DCMs had limb salvage (89%) or mild limb necrosis (11%). Conclusions. 3DCM culture promotes the efficient vascular differentiation of stem cells, and 3DCM transplantation results in the direct vascular regeneration of the injected cells and an improved therapeutic efficacy.