PDF(Pubmed)
Abstract：
Cellular heterogeneity represents a major challenge for regenerative treatment using freshly isolated Adipose Derived Regenerative Cells (ADRCs). Emerging data suggest superior efficacy of ADRCs as compared to the ex vivo expanded and more homogeneous ADRCs (= ASCs) for indications involving (micro)vascular deficiency, however, it remains unknown which ADRC cell subtypes account for the improvement. Surprisingly, we found regarding erectile dysfunction (ED) that the number of injected CD31+ ADRCs correlated positively with erectile function 12 months after one bolus of autologous ADRCs. Comprehensive in vitro and ex vivo analyses confirmed superior pro-angiogenic and paracrine effects of human CD31+ enriched ADRCs compared to the corresponding CD31- and parent ADRCs. When CD31+, CD31- and ADRCs were co-cultured in aortic ring- and corpus cavernous tube formation assays, the CD31+ ADRCs induced significantly higher tube development. This effect was corroborated using conditioned medium (CM), while quantitative mass spectrometric analysis suggested that this is likely explained by secretory pro-angiogenic proteins including DKK3, ANGPT2, ANAX2 and VIM, all enriched in CD31+ ADRC CM. Single-cell RNA sequencing showed that transcripts of the upregulated and secreted proteins were present in 9 endothelial ADRC subsets including endothelial progenitor cells in the heterogenous non-cultured ADRCs. Our data suggest that the vascular benefit of using ADRCs in regenerative medicine is dictated by CD31+ ADRCs.
摘要：
细胞异质性代表了使用新鲜分离的脂肪来源再生细胞（ADRC）进行再生治疗的主要挑战。新的数据表明，与离体扩增和更均匀的ADRCs（=ASC）相比，ADRCs对于涉及（微）血管缺陷的适应症的疗效更好。然而,尚不清楚哪种ADRC细胞亚型导致了这种改善。令人惊讶的是,我们发现,关于勃起功能障碍(ED),注射一次自体ADRCs后12个月,注射CD31+ADRCs的数量与勃起功能呈正相关.全面的体外和离体分析证实，与相应的CD31和亲本ADRC相比，富含人CD31的ADRC具有优异的促血管生成和旁分泌作用。当CD31+时,CD31-和ADRC在主动脉环和海绵体形成试验中共培养，CD31+ADRCs显著诱导更高的管发育。使用条件培养基(CM)证实了这一效应，虽然定量质谱分析表明，这可能是由分泌性促血管生成蛋白（包括DKK3，ANGPT2，ANAX2和VIM）解释的，全部富含CD31+ADRCCM。单细胞RNA测序显示上调和分泌蛋白的转录本存在于9个内皮ADRC亚群中，包括异源非培养ADRC中的内皮祖细胞。我们的数据表明，在再生医学中使用ADRC的血管益处由CD31ADRC决定。
