Journal: Frontiers in Cell and Developmental Biology

Article Title: Morphological and Molecular Features of Porcine Mesenchymal Stem Cells Derived From Different Types of Synovial Membrane, and Genetic Background of Cell Donors

doi: 10.3389/fcell.2020.601212

Figure Lengend Snippet: SMSCs derived from different synovial tissue sources in DL and AS breeds successfully differentiated into adipocytes, osteocytes, and chondrocytes under optimal conditions. In the late stages of osteogenesis after culturing in an osteogenic medium for 28 days, calcium deposits were revealed as brown–black lines or spots under a 4× objective phase-contrast microscope and as red-brown after being stained with alizarin red. The lipid-vacuole and lipid-droplet formation of adipocytes was observable at 4 days of adipogenic differentiation; the cells were fixed and stained with Oil Red O to identify the lipid vacuoles (red). For chondrogenic differentiation, the cells were cultivated in a chondrogenic medium for 35 days; sulfated proteoglycans, including hyaluronic acid, were then stained with alcian blue 8GX (bluish-green/blue). The control cultured un-differentiated cells had a fibroblastic morphology under phase contrast similar to that observed before differentiation to chondrocytes, adipocytes, and osteocytes.

Article Snippet: The adipogenic induction medium (StemPro TM adipogenesis differentiation kit) and chondrogenic induction medium (StemPro TM chondrogenesis differentiation Kit) were purchased from Thermofisher.

Techniques: Derivative Assay, Microscopy, Staining, Control, Cell Culture

Journal: Stem Cells International

Article Title: Is There a Noninvasive Source of MSCs Isolated with GMP Methods with Better Osteogenic Potential?

doi: 10.1155/2019/7951696

Figure Lengend Snippet: Characterization of the profile of DPSCs, OOMDSCs, and UC-MSCs.

Article Snippet: After 24 hours of culture in a basal culture medium, the culture medium was changed to the specific chondrogenic differentiation medium supplemented with growth factors (StemPro® Chondrogenic Differentiation Kit; Gibco Invitrogen, Grand Island, NY).

Techniques: In Vitro, Marker, Standard Deviation

Journal: Stem Cells International

Article Title: Is There a Noninvasive Source of MSCs Isolated with GMP Methods with Better Osteogenic Potential?

doi: 10.1155/2019/7951696

Figure Lengend Snippet: Multilineage differentiation in vitro. Row A: OOMDSC; row B: DPSC; and row C: UC-MSC. (a) The control group of undifferentiated strains. (b) Adipogenic differentiation after eighteen days of induction and staining with oil red; white arrows show the fat vesicles. (c) Chondrogenic differentiation after 3 weeks of induction, stained with alcian blue; white arrows show the extracellular matrix formation—mucopolysaccharides. (d) Osteogenic differentiation after 3 weeks of OOMDSC induction, stained with alizarin red S; white arrows show the extracellular matrix deposition.

Article Snippet: After 24 hours of culture in a basal culture medium, the culture medium was changed to the specific chondrogenic differentiation medium supplemented with growth factors (StemPro® Chondrogenic Differentiation Kit; Gibco Invitrogen, Grand Island, NY).

Techniques: In Vitro, Staining

Journal: Bioinformation

Article Title: Evaluation of in vitro chondrocytic differentiation: A stem cell research initiative at the King Abdulaziz University, Kingdom of Saudi Arabia

doi: 10.6026/97320630014053

Figure Lengend Snippet: Toluidine blue histology. (A) hWJSCs cultured in chondrogenic basal medium for 21 days, showing normal spindle shaped cells (B) hWJSCs cultured in chondrogenic medium with supplements for 21 days showing positive staining and rounded chondrocyte like cells as indicated by black arrows. (Magnification 10X).

Article Snippet: The hWJSCs (2 x104 cells/well) were plated in a 24 well tissue culture plate and exposed to StemPro chondrogenic differentiation basal media (A100071-01, Thermo Fisher Scientific) fortified with chondrogenic supplements (StemPro Kit content) for up to 21 days with fresh changes of media every 72h.

Techniques: Cell Culture, Staining

Journal: Bioinformation

Article Title: Evaluation of in vitro chondrocytic differentiation: A stem cell research initiative at the King Abdulaziz University, Kingdom of Saudi Arabia

doi: 10.6026/97320630014053

Figure Lengend Snippet: Quantitative real time gene expression analysis of the cartilage related genes in hWJSCs cultured in chondrogenic media for 21 days. Increased expression of (A) collagen, type II, alpha 1 [COL2A1], (B) aggrecan [ACAN] and (C) SRY (sex determining region Y)-box 9 [SOX9] was observed compared to controls. The values were expressed as mean ± SEM from 3 experimental replicates. Asterisk (*) indicate statistical significance at p<0.05 compared to control.

Article Snippet: The hWJSCs (2 x104 cells/well) were plated in a 24 well tissue culture plate and exposed to StemPro chondrogenic differentiation basal media (A100071-01, Thermo Fisher Scientific) fortified with chondrogenic supplements (StemPro Kit content) for up to 21 days with fresh changes of media every 72h.

Techniques: Gene Expression, Cell Culture, Expressing, Control

Journal: eBioMedicine

Article Title: A personalized medicine approach identifies enasidenib as an efficient treatment for IDH2 mutant chondrosarcoma

doi: 10.1016/j.ebiom.2024.105090

Figure Lengend Snippet: Effect of enasidenib treatment in the chondrogenic differentiation pathway and proliferative status. (a–c) Heatmap plots depicting the expression values of genes belonging to the GO BP chondrocyte differentiation pathway according to enasidenib treatment for each cell line. (b–c) Histological analysis of formalin-fixed paraffin-embedded T-CDS17#1 cell spheroids growth in chondrocyte differentiation medium with or without (control) 10 μM enasidenib for 21 days. Representative images of H&E and PAS-alcian staining (b) and quantification of PAS-alcian stain (c) are displayed. Scale bars = 100 μm. Error bars represent the standard deviation of three independent experiments. (d) Heatmap plot showing the expression values of those genes belonging to the mRNA expression signature developed by Nicolle et al. (2019), including their microarray experiments and the classification of chondrosarcoma samples into E1/E2 subtypes (n = 102). (e) Bubble plots showing the top 5 most significant pathways (ORA, FDR <0.05) from the MSigDB GO BP collection in each cell type for up-regulated (left) and down-regulated (right) E1/E2 genes.

Article Snippet: The pellet was cultured in 500 μl of either standard culture medium or ready-to-use complete chondrogenic differentiation medium StemPro chondrogenesis differentiation kit (Cat#A1007101, Gibco-Thermo Fisher, Waltham, MA, USA) following the manufacturer's recommendations.

Techniques: Expressing, Formalin-fixed Paraffin-Embedded, Control, Staining, Standard Deviation, Microarray

Journal: Life

Article Title: Does Needle Design Affect the Regenerative Potential of Bone Marrow Aspirate? An In Vitro Study

doi: 10.3390/life11080748

Figure Lengend Snippet: Differentiation into the three lineages. Exemplary presentation of osteogenic, adipogenic and chondrogenic (left to right) differentiation, each with the respective control without differentiation medium. The small black background bar represents 200 µm.

Article Snippet: For chondrogenic differentiation, a micromass culture with 800,000 cells was seeded in 96-round bottom wells (Greiner) corresponding to a cell density of 22,860 cells/cm 2 in chondrogenic differentiation medium (StemPro® Osteocyte/Chondrocyte Differentiation Basal Medium/Chondrogenesis Supplement, Fisher Scientific).

Techniques: Control

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: Primers designed for real-time PCR and the respective amplicon sizes of the products

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques: Real-time Polymerase Chain Reaction, Amplification, Marker

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: Growth characteristics of DPSCs and aBMMSCs after long-term expansion with three different culture media: one serum-based (CCM) and two serum/xeno-free, cGMP media (StemMacs and StemPro). a Population doubling (PD) time (days) (mean ± SD) for DPSCs ( n = 6 donors, experiments repeated three times in duplicates) and aBMMSCs ( n = 4 donors, experiments repeated three times in duplicates) through passages. Comparisons between cells, passages and media performed by two-way ANOVA followed by Tukey’s post-hoc tests. Asterisks indicate statistically significant increase in PDT at consecutive passages of each culture medium/cell type (* p < 0.05; ** p < 0.01). b Total cumulative PD numbers (mean ± SD) for DPSCs ( n = 6) and aBMMSCs ( n = 4) calculated based on the ratio of cells seeded vs cells harvested per passage. Asterisks indicate statistically significant differences (* p < 0.05; ** p < 0.01; n.s. = nonsignificant, calculated by two-way ANOVA followed by Tukey’s post-hoc tests) either among the three different media (depicted below/above red or blue double arrows for DPSCs and aBMMSCs, respectively) or between the two different cell types (over horizontal black double arrows). aBMMSC alveolar bone marrow mesenchymal stem cell, CCM complete culture medium, DPSC dental pulp stem cell, P cell passage

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques:

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: Morphological characteristics of DPSCs and aBMMSCs after long-term expansion with three different culture media: one serum-based (CCM) and two serum/xeno-free, cGMP media (StemMacs and StemPro). a, b Phase-contrast microscopy photographs of DPSCs and aBMMSCs, respectively (sale bars: 100 μm). c, d Flow cytometry fluorescence intensity plots of forward scatter (FSC) vs side scatter (SSC) parameters corresponding to the cell size and cell internal complexity (granularity), respectively. aBMMSC alveolar bone marrow mesenchymal stem cell, CCM complete culture medium, DPSC dental pulp stem cell, P cell passage

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques: Microscopy, Flow Cytometry, Fluorescence

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: Impact of long-term expansion of DPSCs and aBMMSCs using three different culture media (CCM, StemMacs and StemPro) on β-galactosidase activity. a, b Optical microscopy photographs of DPSCs and aBMMSCs, respectively (sale bars: 100 μm). c Percentage of SA-β-gal-positive cells (DPSCs and aBMMSCs) at early, middle and late passages of each expansion medium. Values are mean (± SD) of DPSCs ( n = 6 donors, experiments repeated three times in duplicates) and aBMMSCs ( n = 4 donors, experiments repeated three times in duplicates). Asterisks indicate statistically significant differences (* p < 0.05; ** p < 0.01) between middle vs early and late vs early passages for each cell type and medium separately. Upper case letters (A) indicate statistically significant differences between DPSCs and aBMMSCs expanded with the same medium (either CCM, StemMacs or StemPro) at each passage separately (either early, middle or late), while lowercase letters (a) indicate statistically significant differences between StemMacs vs CCM and between StemPro vs CCM for each cell type (either DPSCs or aBMMSCs) and each passage (either early, middle or late) separately. Statistical analyses were performed by two-way ANOVA followed by Tukey’s post-hoc tests. aBMMSC alveolar bone marrow mesenchymal stem cell, CCM complete culture medium, DPSC dental pulp stem cell, P cell passage, SA-β-gal beta-galactosidase

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques: Activity Assay, Microscopy

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: Impact of long-term expansion of DPSCs and aBMMSCs with three different culture media (CCM, StemMacs and StemPro) on telomere length. Telomere length determined by Southern blot analysis in cells harvested at early (p.2–3), middle (p.6–7) or late (p.10–11) passages. Lane 2 serves as a technical control, to confirm that the hybridization and Southern blot procedures were successful. Lanes 1 and 12 are molecular weight markers (kbp). a, c Representative Southern blots and b, d respective mean terminal restriction fragments (TRFs ± SD) for each medium/passage. A trend for lower mean TRF values could be observed with continuous passaging. This was consistent for all different culturing media and types of cells. Figures are representative of a single DPSC and aBMMSC donor for each culture medium. aBMMSC alveolar bone marrow mesenchymal stem cell, CCM complete culture medium, DPSC dental pulp stem cell, p. cell passage

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques: Southern Blot, Hybridization, Molecular Weight, Passaging

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: a–l Real-time PCR analysis of the expression of lineage-specific osteogenic (ALP, BMP-2), chondrogenic (SOX-9, ACAN) and adipogenic markers (PPAR-γ, LPL) in DPSCs ( n = 3 donors, D1–D3) and aBMMSCs ( n = 3 donors, D1–D3) after long-term expansion with three different culture media (CCM, StemMacs and StemPro). Values are mean (± SD) of three independent experiments ( n = 3) in duplicate. Asterisks indicate statistically significant differences (* p < 0.05; ** p < 0.01) in expression of each marker with passaging for each cell type/donor and medium separately. Comparisons between cells, passages and media were performed by two-way ANOVA followed by Tukey’s post-hoc tests. aBMMSC alveolar bone marrow mesenchymal stem cell, ACAN aggrecan, ALP alkaline phosphatase, BMP-2 bone morphogenetic protein, CCM complete culture medium, DPSC dental pulp stem cell, LPL lipoprotein lipase, P cell passage, D donor, PPAR-γ peroxisome proliferator-activated receptor gamma, SOX-9 sex-determining region Y-Box 9

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Marker, Passaging

Journal: Stem Cell Research & Therapy

Article Title: Isolation and prolonged expansion of oral mesenchymal stem cells under clinical-grade, GMP-compliant conditions differentially affects “stemness” properties

doi: 10.1186/s13287-017-0705-0

Figure Lengend Snippet: Real-time PCR analysis of the expression of osteogenic (ALP, BMP-2, BGLAP) and chondrogenic (SOX-9, ACAN) markers after induction of early, middle and late-passage DPSCs and aBMMSCs expanded previously with three different culture media (CCM, StemMacs and StemPro). Data are representative of one cell donor for each cell type and medium. Values are mean (± SD) of three independent experiments ( n = 3) in duplicate. Asterisks over horizontal double arrows indicate statistically significant differences (* p < 0.05; ** p < 0.01; n.s. = nonsignificant) at each passage of each cell type and medium, during the entire induction period (D0, D7 and D14). Asterisks over/under red, blue or green brackets indicate statistically significant differences (* p < 0.05; ** p < 0.01) in the expression of each marker among different passages (p.2 vs p.6 vs p.10) at D14 after induction of differentiation. a–f Expression of osteogenic markers (ALP, BMP-2, BGLAP). g , h Spectrophotometric quantification of the Alizarin Red S staining of DPSCs and aBMMSCs induced for osteogenic differentiation after 21 days (mean nmol AR-S/μg of total protein ± SD of three independent experiments of a representative donor in three replicates). i–l Expression of chondrogenic markers (SOX-9, ACAN) at D0, D7 and D14 after induction. Comparisons between cells, passages and media performed by two-way ANOVA followed by Tukey’s post-hoc tests. aBMMSC alveolar bone marrow mesenchymal stem cell, ACAN aggrecan, ALP alkaline phosphatase, BGLAP bone gamma-carboxyglutamate protein, BMP-2 bone morphogenetic protein, CCM complete culture medium, DPSC dental pulp stem cell, P cell passage, D day, SOX-9 sex-determining region Y-Box 9

Article Snippet: After 2–3 days, confluent MSCs were exposed to either osteogenic or chondrogenic media according to the manufacturer’s instructions (StemPro Osteogenic and StemPro Chondrogenic Differentiation Kits; Life Technologies).

Techniques: Real-time Polymerase Chain Reaction, Expressing, Marker, Staining

Journal: PLoS ONE

Article Title: Peripheral Blood Mononuclear Cells Enhance Cartilage Repair in in vivo Osteochondral Defect Model

doi: 10.1371/journal.pone.0133937

Figure Lengend Snippet: Morphological analysis of the adherent PBMCs (A-F) and Mesoblast MSCs (G-L) in both normoxic and hypoxic culture at day 21 under phase contrast light microscopy. Representative images of osteogenic differentiation (Alizarin-red; A, D, G and J), adipogenic differentiation (Oil-red-O; B, E, H and K) and chondrogenic differentiation (Alcian blue; C, F, I and L). Scale bar 200 μm.

Article Snippet: To promote chondrogenic differentiation StemPro Chondrogenesis Supplement (A10064-10, Invitrogen) was added to the basal medium and the medium was changed every 3–4 days for 21 days [ ].

Techniques: Light Microscopy