Journal: Journal of visualized experiments : JoVE

Article Title: Cell Dissociation from the Tongue Epithelium and Mesenchyme/Connective Tissue of Embryonic-Day 12.5 and 8-Week-Old Mice

doi: 10.3791/62163

Figure Lengend Snippet: Materials

Article Snippet: Name Company Catalog Number Comments bovine serum albumin (BSA) Gold Biotechnology A-420-100 C57BL/6 mouse (C57BL/6J) The Jackson Laboratory 000664 collagenase (Collagenase A) Sigma-Aldrich 10103586001 culture dish (35 mm in diameter) Genesee Scientific 32-103G culture dish (100 mm in diameter) Genesee Scientific 32-107G dispase (Dispase II) Sigma-Aldrich 04942078001 dissecting scissors (Student Fine Scissors) Find Science Tool 91460-11 DMEM/F12 Gibco 11320033 fetal bovine serum (FBS) Hyclone C838U82 fine forceps (Dumount #3 Forceps) Find Science Tool 11293-00 hemocytometer Hausser Scientific 3520 inverted microscope with imaging system (EVOS XL Core Cell Imaging System) Life Technologies AMEX1000 low retention pipette tips METTLER TOLEDO 17014342 mini-scissors (Evo Spring Scissors) Fine Science Tool 15800-01 plastic warp VWR 46610-056 spatula (Moria Spoon) Fine Science Tool 10321-08 surgical forceps (Dumount #2 Laminectomy Forceps) Fine Science Tool 11223-20 Trypan blue Gibco 15250061 Tyrode’s solution Sigma-Aldrich T2145-10L made from Tyrode’s salts 0.25% typsin-EDTA Gibco 25200056 0.1 M Phosphate-Buffered Saline (PBS) Hoefer 33946 made from 1 M PBS 0.22-μm syringe filter Genesee Scientific 25-243 70% ethanol Koptec 233919 made from 100% ethanol 1-mL syringe BD 8194938 5-mL low binding microcentrifuge tube Eppendorf 30122348 30-G needle BD 9193532 35-μm cell strainer Falcon 64750 70-μm cell strainer Falcon 64752 Open in a separate window Materials.

Techniques: Inverted Microscopy, Imaging, Transferring, Saline, Binding Assay

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cross-Tissue Characterization of Heterogeneities of Mesenchymal Stem Cells and Their Differentiation Potentials

doi: 10.3389/fcell.2021.781021

Figure Lengend Snippet: MSC subpopulations in 4 different tissues. (A) . UMAP projection of MSCs in 4 different tissues, colored by MSC subpopulations. (B) . UMAP projection of MSCs from each tissue, namely UC-MSCs, BM-MSCs, SY-MSCs, and AD-MSCs. (C) . Dot plot of the expression of subpopulation-specific genes in the MSC subpopulations. (D) . Heatmap of AUROC scores between MSC subpopulations pairs, in which AUROC scores represent similarities between subpopulations. (E) . Weighted ligand-receptor interactions between MSC subpopulations pairs in each tissue based on STRING database. The edge thickness and corresponding numbers indicate the sum of weighted paths between subpopulations.

Article Snippet: The Wharton’s jelly was cut into small pieces (2–3 mm 3 ), which were placed in a petri dish with MSCs culture medium (MesenGro ®human MSC Medium, StemRD, US), 10% fetal bovine serum (FBS; Gibco, Australia), and 10 μg/L basic fibroblast growth factor (bFGF; Gibco, Australia).

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cross-Tissue Characterization of Heterogeneities of Mesenchymal Stem Cells and Their Differentiation Potentials

doi: 10.3389/fcell.2021.781021

Figure Lengend Snippet: Features and lineage-specific differentiation potentials of MSC subpopulations. (A) . Violin plot of stemness scores of MSCs in 4 different tissues. (B) . Bar plot of the expression level of AMIGO2, CLDN1, LRRC17, and SLC22A3 in each cell. (C) . Violin plot of immunoregulatory scores of MSCs in 4 different tissues. (D) . Bar plot of the expression level of AREG, CSF3, CCL20, and IL6 in each cell. (E–G) . Violin plot of osteocyte score. (E) , adipocyte score. (F) , and chondrocyte score. (G) of MSCs in MSC subpopulations. (H) . Fractions of MSC subpopulations in MSCs for each tissue.

Article Snippet: The Wharton’s jelly was cut into small pieces (2–3 mm 3 ), which were placed in a petri dish with MSCs culture medium (MesenGro ®human MSC Medium, StemRD, US), 10% fetal bovine serum (FBS; Gibco, Australia), and 10 μg/L basic fibroblast growth factor (bFGF; Gibco, Australia).

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cross-Tissue Characterization of Heterogeneities of Mesenchymal Stem Cells and Their Differentiation Potentials

doi: 10.3389/fcell.2021.781021

Figure Lengend Snippet: Response heterogeneity and dynamics of MSCs to chondrogenesis induction and its underlying mechanisms. (A) . t-SNE projection of MSCs pre-and post- chondrogenesis induction, colored by non-induced UC-MSC (UC-MSCs) and chondrogenic-induced MSCs (UCMSC-I). (B) . t-SNE projection of MSCs pre-and post- chondrogenesis induction, colored by the six MSC subpopulations, namely chondro, osteo, adipo, chondro-I, osteo-I, and adipo-I. (C) . Dot plot of the expression level of lineage-specific genes in MSC subpopulations pre-and post-chondrogenesis induction. (D) . Boxplot of chondrocyte differentiation scores of pre-induced chondro (Chondro) and chondrogenic-induced chondro (Chondro-I). Wilcoxon Rank Sum test, *** p < 0.001. (E) . Fractions of MSC subpopulation in MSCs and chondrogenic-induced MSCs. Fisher’s exact test, * p value <0.01, *** p value <0.001. (F) . The significantly enriched GO terms in chondro-I specific genes compared to non-induced chondro. (G) . Heatmap of the most significantly differentially expressed genes between pro-and post-induced chondro. (H) . Bar plot of significant interactions between MSC subpopulations pairs in UC-MSCs and UCMSC-I. (I) . Bubble plot of FOSL2 and its target genes, with each target gene colored by log fold change between post-induced chondro and pre-induced chondro.

Article Snippet: The Wharton’s jelly was cut into small pieces (2–3 mm 3 ), which were placed in a petri dish with MSCs culture medium (MesenGro ®human MSC Medium, StemRD, US), 10% fetal bovine serum (FBS; Gibco, Australia), and 10 μg/L basic fibroblast growth factor (bFGF; Gibco, Australia).

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cross-Tissue Characterization of Heterogeneities of Mesenchymal Stem Cells and Their Differentiation Potentials

doi: 10.3389/fcell.2021.781021

Figure Lengend Snippet: Response heterogeneity and dynamics of MSCs to osteogenesis induction and its underlying mechanisms. (A) . t-SNE projection of MSCs pre-and post-osteogenesis induction, colored by non-induced BM-MSC (BM-MSCs) and osteogenic-induced MSCs (BMMSC-I). (B) . t-SNE projection of MSCs pre-and post-osteogenesis induction, colored by the six MSC subpopulations, namely chondro, osteo, adipo, chondro-I, osteo-I, and adipo-I. (C) . Dot plot of the expression level of lineage-specific genes in MSC subpopulations pre- and post-osteogenesis induction. (D) . Boxplot of osteocyte differentiation scores of cells in BMMSC and post- induced osteo (osteo-I). Wilcoxon Rank Sum test, *** p < 0.001. (E) . Cell fractions of MSC subpopulation in BMMSCs and BMMSC-I. Fisher’s exact test, * p -value < 0.05. (F) . The significantly enriched GO terms in osteo-I specific genes compared to non-induced osteo. (G) . Heatmap of differentially expressed genes between pre- and post-induced osteo. (H) . Bar plot of significant interaction between MSC subpopulations pairs in BM-MSCs and BMMSC-I. (I) . FOXO3 and its target genes, with each target gene colored by log fold change between osteogenic-induced osteo and pre-induced osteo.

Article Snippet: The Wharton’s jelly was cut into small pieces (2–3 mm 3 ), which were placed in a petri dish with MSCs culture medium (MesenGro ®human MSC Medium, StemRD, US), 10% fetal bovine serum (FBS; Gibco, Australia), and 10 μg/L basic fibroblast growth factor (bFGF; Gibco, Australia).

Techniques: Expressing

Journal: Frontiers in Cell and Developmental Biology

Article Title: Cross-Tissue Characterization of Heterogeneities of Mesenchymal Stem Cells and Their Differentiation Potentials

doi: 10.3389/fcell.2021.781021

Figure Lengend Snippet: Response heterogeneity and dynamics of MSCs to adipogenesis induction and its underlying mechanisms. (A) . t-SNE projection of MSCs pre-and post-adipogenesis induction, colored by non-induced BM-MSC (BM-MSCs) and adipogenic-induced MSCs (BMMSC-I). (B) . t-SNE projection of MSCs pre-and post-adipogenesis induction, colored by MSC subpopulations, namely chondro, osteo, adipo, myo, chondro-I, osteo-I, adipo-I, and myo-I. (C) . Fractions of MSC subpopulation in BM-MSCs and adipogenesis-induced MSCs. Fisher’s exact test, * p -value < 0.05. (D) . Significantly enriched GO terms in adipo-specific genes. (E) . Expression of adipogenic lineage-specific genes on t-SNE projection of MSCs. (F) . Expression of CEBPA in single cells (left panel) , CEBPA and its target genes, with each target gene colored by log fold change between adipogenesis-induced adipo and pre-induced adipo (right panel) . (G) . Expression of PPARG in single cells (left panel) , PPARG and its target genes, with each target gene colored by log fold change between adipogenesis-induced adipo and pre-induced adipo (right panel) .

Article Snippet: The Wharton’s jelly was cut into small pieces (2–3 mm 3 ), which were placed in a petri dish with MSCs culture medium (MesenGro ®human MSC Medium, StemRD, US), 10% fetal bovine serum (FBS; Gibco, Australia), and 10 μg/L basic fibroblast growth factor (bFGF; Gibco, Australia).

Techniques: Expressing