Pre-culture in endothelial growth medium enhances the angiogenic properties of adipose-derived stem/stromal cells

Abstract

Considerable progress has been made on the development of adipose-derived stem/stromal cells (ASCs) as pro-angiogenic therapeutic tools. However, variable clinical results highlight the need for devising strategies to enhance their therapeutic efficacy. Since ASCs proliferate and stabilize newly formed vessels during the angiogenic phase of adipose tissue formation, we hypothesized that mimicking an angiogenic milieu during culture of ASCs would enhance their capacity to support endothelial cell survival and angiogenesis. To test this, we compared the effect of an endothelial growth medium (EGM-2) and conventional media (αMEM) on the progenitor and angiogenic properties of ASCs. ASCs cultured in EGM-2 (ASC-EGM) displayed the highest clonogenic efficiency, proliferative potential and multilineage potential. After co-culture under growth factor starvation, only ASC-EGM attenuated luciferase-expressing human umbilical vein endothelial cells (HUVEC) apoptosis and supported the formation of endothelial cords in a dose-dependent manner. These effects were recapitulated by the conditioned medium of ASC-EGM, which displayed a 100-fold higher expression of hepatocyte growth factor in comparison with ASC-αMEM. Next, HUVEC and ASCs were co-transplanted subcutaneously into immunodeficient mice, and the survival of HUVEC was monitored by bioluminescent imaging. After 60 days, the survival of HUVEC transplanted alone was equivalent to that of HUVEC co-transplanted with ASC-αMEM (15.0 ± 0.7 vs. 13.0 ± 0.5%). Strikingly, co-transplantation with ASC-EGM increased HUVEC survival to 105.0 ± 3.5%, and the resulting organoids displayed functional vasculature with the highest human-derived vascular area. These findings demonstrate that pre-conditioning of ASCs in endothelial growth medium augment their pro-angiogenic properties and could enhance their therapeutic efficacy against ischemic diseases.

RETRACTED ARTICLE: Sirtuin 3 inhibition induces mitochondrial stress in tongue cancer by targeting mitochondrial fission and the JNK-Fis1 biological axis

“…In rectal cancer (Li et al 2017), aberrant mitochondrial fission mediates cellular oxidative stress that blunts cancer migration. In the present study, our results indicated that mitochondrial fission was associated with mitochondrial apoptosis and mitochondrial ROS overproduction (Koopman et al 2017;Zhou et al 2017d); inhibition of mitochondrial fission sustained mitochondrial metabolism and attenuated mitochondrial damage, finally promoting cancer cell survival (Souza et al 2018). Therefore, our results combined with those of previous studies lay the foundation for understanding the molecular features of mitochondrial fission in mitochondrial damage and substantiate the sufficiency and necessity of mitochondrial fission in inducing cancer death (Koentges et al 2017;Reddy et al 2018).…”

High‐throughput fabrication of vascularized adipose microtissues for 3D bioprinting

“…Capillary‐like networks are defined as a homogeneous web or dense region of endothelial cells. The VAT‐2S ASC/HUVEC spheroids co‐cultured in EGM presented capillary‐like structures, in line with previous studies, showing that the endothelial cells were stimulated to proliferate and formed capillary‐like structures (Borges et al, 2003; Frerich et al, 2008; Kang et al, 2009; Souza et al, 2018). Conversely, the VAT‐2S spheroids co‐cultured in 1:1 EGM/ADM showed only successful adipogenic differentiation but did not present prevascular structures.…”

Effects of VEGF‐ANG‐1‐PLA nano‐sustained release microspheres on proliferation and differentiation of ADSCs

“…Our research emphasis was that the underlying role of VEGF‐Ang‐1‐PLA nano‐sustained release microspheres on ADSC proliferation and differentiation toward ECs. It was reported that pre‐culture in endothelial growth medium enhanced the angiogenic properties of ADSCs (Souza et al, ), and Sophia Khan found that the two vital angiogenesis factors FGF and VEGF played critical roles in endotheliogenesis from human ADSCs (Khan et al, ). The present study VEGF‐Ang‐1‐PLA microspheres facilitated HGF secretion to promote ADSC proliferation and enhance CD31 + cells, but scarcely influenced the expression levels of VEGF and Ang‐1.…”