Adipose-derived dual cell therapy enhances arteriogenesis and limb preservation through vascular integration in critical limb ischemia

Peripheral artery disease (PAD) causes progressive arterial narrowing in the lower limbs and can advance to critical limb ischemia (CLI). Limited revascularization options highlight the need for safer, more effective therapies. Vascular multipotent stem cells (VMSCs) and adipose-derived stem cells (ADSCs) were isolated from adipose tissue, characterized phenotypically, and tested for angiogenic activity in vitro. Their therapeutic efficacy was then examined in a murine critical limb ischemia model through intramuscular transplantation, assessing limb preservation, neovascularization, and cell integration. VMSCs shared mesenchymal stem cell-like features with ADSCs and exhibited robust proliferative capacity, enabling rapid expansion to clinically relevant numbers. VMSCs also demonstrated endothelial-like properties, including CD31, VE-cadherin, and CD141 expression, and formed capillary-like structures in vitro. In contrast, ADSCs displayed perivascular characteristics with α-SMA and Transgelin expression. Co-culture of VMSCs and ADSCs promoted the development of mature tubular networks in vitro. Combined cell transplantation markedly decreased limb loss and promoted both angiogenesis and arteriogenesis in ischemic tissue, with transplanted cells partially integrating into the host vasculature to form hybrid vascular structures. VMSCs and ADSCs show complementary regenerative functions, sustained engraftment, and support for large-vessel formation, underscoring their potential for stem cell-based vascular therapies.