Strain C57BL/6 Mouse Adipose-Derived Mesenchymal Stem Cells with GFP

Single-Cell Transcriptomics of mADSCs-Treated and Untreated Pulmonary Resident Cells

Idiopathic pulmonary fibrosis (IPF) is a severe lung disease with few effective treatments. ADSCs have demonstrated potential benefits for IPF, but the underlying mechanisms are unknown. In this study, bleomycin (BLM) was used to induce pulmonary fibrosis in mice, after which GFP-labeled mouse adipose-derived mesenchymal stem cells (mADSCs) were intratracheally implanted. The mADSCs were retrieved and analyzed via single-cell RNA sequencing to investigate their impact on the macrophage profile and overall lung microenvironment.

Firstly, Rahman et al. analyzed single-cell profiles of lung parenchyma cells in BLM + PBS and BLM + mADSCs groups (Fig. 1b), containing 8181 and 6405 cells respectively. Both groups revealed 27 cell clusters, and 12 cell types were identified based on markers, without cell cycle impact (Fig. 1b). The lung cell types included pneumocytes, bronchiolar exocrine cells, natural killer cells, fibroblasts/epithelial cells, myofibroblasts, dendritic cells, endothelial cells, T cells, B cells, monocytes/macrophages, and granulocytes (Fig. 1b, c). Key findings showed monocytes/macrophages (48.78%) in the BLM + mADSCs group, and granulocytes (44.87%) in the BLM + PBS group as prevalent immune cells (Fig. 2d). The primary non-immune cells in BLM + mADSCs were fibroblasts, endothelial cells, and myofibroblasts, occupying 7.45, 6.83, and 3.80% respectively. Monocytes/macrophages were also predominant in both groups BLM + mADSCs (48.78%) and BLM + PBS (27.28%) (Fig. 2a), split into 9 subclasses based on gene expression (Fig. 2b). Mo/Ma-1, marked by MPG, COL1a1, and SPARC, was the largest subgroup (48.71%), whereas macrophage-4 expressing GPNMB, PF4, and APOE was the second largest in BLM + mADSCs (20.42%). Alveolar macrophage-21 and Mo/Ma-24 were negligible in BLM + PBS (0 and 0.07%) (Fig. 2b). Further analysis on Mo/Ma-1 and macrophage-4 subclasses showed they impact ECM regulation and related pathways with genes like COL1a1, COL1a2, and SPARC highly expressed (Fig. 2c, d).

Fig. 1. Injection of mADSCs through the intratracheal caused increase of monocyte/macrophage population (Rahman M, Wang Z Y, et al., 2021).

Fig. 2. Go enrichment and KEGG analysis of monocyte/macrophage (a) UMAP distribution of 9 monocyte/macrophage clusters in both groups (Rahman M, Wang Z Y, et al., 2021).

Adipose-Derived Mesenchymal Stem Cell Seeded Atelocollagen Scaffolds for Cardiac Tissue Engineering

Stem cell transplantation presents a possible treatment option for myocardial infarction through the engineering of heart tissue with diverse biological materials. Research indicates that Atelocollagen displays promising scaffold properties because of its ability to function well with biological tissues. Adipose-derived mesenchymal stem cell (ADMSCs) can be readily extracted from their source and show promise in tissue engineering applications.

Li's team explored the biocompatibility of mouse ADMSCs-GFP with Atelocollagen scaffolds and their ability to differentiate into cardiomyocytes for myocardial tissue engineering. Proliferation of ADMSCs on the collagen scaffolds of Atelocollagen was evaluated by measuring the fluorescence of GFP labeled ADMSCs. The ADMSCs grew rapidly on the collagen scaffolds of Atelocollagen, and the cells can grow into the mesh of the collagen scaffolds. On the 7th day, about 50% of the mesh of the scaffolds were filled with GFP-labeled ADMSCs (Fig. 3). ADMSCs were cultured on 3D Atelocollagen scaffolds. After 3 weeks of 5-aza induction, cTnT immunofluorescence was positive (Fig. 4).

Fig. 3. The image of ADMSCs compatible with the Atelocollagen scaffolds (Li Q, Li MM, et al., 2020).

Fig. 4. cTnT expression of ADMSCs induced 3 weeks with 5-aza on 3D scaffolds by immunofluorescence staining (Li Q, Li MM, et al., 2020).