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ScRNA-seq profiling of vascular cells isolated from disease-free human breast based on the expression of CD36 and/or CD31

ABSTRACT: To identify markers preferentially expressed by capillary ECs (highly expressing CD36) in the disease-free human breast and gain insights into their biology, we used single cell RNA sequencing (scRNA-seq). For this analysis, dissociated cells from disease-free tissues were barcoded using the MULTI-seq protocol and pooled prior to FACS-based isolation of CD36+ and/or CD31+ cells. The droplet-based 10x Genomics Chromium platform was used to encapsulate single cells and construct cDNA libraries. Populations of fibroblasts, perivascular, lymphatic and vascular endothelial ECs where identified in this analysis.

ORGANISM(S): Homo sapiens

PROVIDER: GSE190592 | GEO | 2021/12/11

REPOSITORIES: GEO

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Project description:A major challenge in realizing regenerative treatment using freshly isolated Adipose Derived Regenerative Cells (ADRCs) is the cellular heterogeneity and donor variability. Ex vivo expanded ADRCs (=ASCs) are available at larger quantities and represent a more homogeneous population suitable for allogenic use. Emerging pre-clinical and clinical data however suggest similar, or even superior efficacy of ADRCs as compared to ASCs for indications involving (micro)vascular deficiency. Since CD31+ ADRCs lack in cultured ASCs, we hypothesized that these cells account for improvement of vascular function in the heterogenous ADRCs. Herein, we found that in patients with erectile dysfunction (ED), the number of injected CD31+ ADRCs correlates 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. CD31+, CD31- and ADRCs were co-cultured in aortic ring-, as well as in ED-relevant corpus cavernousum tube formation assays, both showing a significantly higher ability of the CD31+ ADRCs to support tube development. This effect was corroborated using conditioned medium (CM), while quantitative mass spectrometric analysis suggested that this is explained by secretory pro-angiogenic proteins such as DKK3, ANGPT2, ANAX2 and VIM, all being enriched in CD31+ADRC CM. To gain further specification of the ADRC subpopulations expressing these proteins, single-cell RNA sequencing was performed and 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 thus suggest that the vascular benefit of using ADRCs in regenerative medicine is dictated by CD31+ ADRCs, which likely represent an improved alternative to heterogenous ADRCs as well as ASCs when aiming for vascular repair in cell therapy.

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ScRNA-seq profiling of vascular cells isolated from disease-free human breast based on the expression of CD36 and/or CD31

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