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Single cell RNA sequencing analysis of native fat tissue, RMF pallets, and expanded ADSCs

ABSTRACT: Objectives: 1. To explore cell population changes after a 4-week in vitro self-reaggregation culture, comparing RMF pellets, native fat tissue, and expanded ADSCs. 2. To analysis the cellular composition and maturity of RMF-islet organoids before and post transplantation. Methods: Sc-Seq was performed on cells from RMF pellets (n = 3), native fat (n = 3), expanded ADSCs (n = 3), in vitro RMF-islet organoids (n = 2), and in vivo RMF-islet organoid grafts (n = 2) using the 10× Genomics platform. Results: A total of 65501 cells were clustered from the RMF pellets, native fat, and expanded ADSC samples and visualized using UMAP. Cells were categorized into main cell types based on their spatial distribution on the UMAP plots: adipose-derived stem and progenitor cell (ASPC) clusters (ASPC-1, ASPC-2, ASPC-3, and ASPC-4), preadipocyte cluster, endothelial cell cluster, smooth muscle cell cluster (SMC), and immune cell cluster. Following a 4-week in vitro culture, the percentage of ASPCs increased from 24.57% in native fat to 97.17% in RMF pellets and further to 99.9% in expanded ADSCs. Pseudotime analyses the ASPC-1 cluster, predominantly derived from native fat samples (96.04%), occupied the initial position on the developmental trajectory, followed by ASPC-2 and ASPC-3 clusters primarily originating from RMF pellet samples (94.91% and 96.55%, respectively). Conversely, the ASPC-4 cluster mainly derived from expanded ADSC samples were positioned at the terminal end of the trajectory. Furthermore, pseudotime values indicated that both ASPC-2 and ASPC-3 clusters from RMF pellet samples exhibited an intermediate differentiation state between those observed in native fat and expanded ADSC samples. For RMF-islet organoids, a total of 8715 cells and 5328 cells were analyzied from the in vitro and in vivo RMF-islet organoid samples, respectively. Compared these data with published transcriptomes of human islet cells, we identified four distinct populations of pancreatic endocrine cells, including β-, α-, δ-, or γ-like cells, within the two in vitro RMF-islet organoid samples and the presence of all four major pancreatic endocrine cell types within the organoid grafts and displayed a similar cellular composition before and after transplantation. Conclusion: 1. ASPC clusters from RMF pellet samples possess an intermediary differentiated phenotype compared to their counterparts derived from native fat and expanded ADSC samples. 2. The RMF-islet organoids harbor a population of functionally matured β-like cells exhibiting transcriptional similarities to native human islet β-cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE280107 | GEO | 2025/10/28

REPOSITORIES: GEO

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Single cell RNA sequencing analysis of native fat tissue, RMF pallets, and expanded ADSCs

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