Journal: Neuron

Article Title: Xenotransplanted Human Cortical Neurons Reveal Species-Specific Development and Functional Integration into Mouse Visual Circuits

doi: 10.1016/j.neuron.2019.10.002

Figure Lengend Snippet:

Article Snippet: On day −2, ESCs were dissociated using Stem-Pro Accutase (Thermo Fisher Scientific, Cat#A1110501) and plated on matrigel- (hES qualified matrigel BD, Cat#354277) coated dishes at low confluency (5,000–10,000 cells/cm2) in MEF-conditioned hES medium supplemented with 10 μM ROCK inhibitor (Y-27632; Merck, Cat#688000).

Techniques: Recombinant, Knock-Out, Transfection, Software

Journal: The Journal of Biological Chemistry

Article Title: Ultraviolet A Regulates Adipogenic Differentiation of Human Adipose Tissue-derived Mesenchymal Stem Cells via Up-regulation of Kruppel-like Factor 2

doi: 10.1074/jbc.M110.135830

Figure Lengend Snippet: UVA irradiation inhibits adipocyte differentiation. Two-day post-confluent hAMSCs (day 0) were treated with the indicated doses of UVA irradiation and then stimulated with STEM PRO® adipocyte differentiation (DIF) media for 3 days. The medium was then replaced with STEM PRO® adipocyte differentiation media every 3 days until the end of the experiment at day 14. The assays were performed on fully differentiated adipocytes (day 14). A, intracellular lipids were stained with Oil Red O. The results were confirmed by three independent experiments, which were each conducted in duplicate. B, triglyceride content was measured using a triglyceride assay kit (Cayman Chemical, Ann Arbor, MI). Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The results were verified by three repetitions of the experiments, each of which was conducted in triplicate. C, GPDH activity was measured using GPDH activity assay kits (Takara Bio Inc., Japan). Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The results were verified by four repetitions of the experiments. D, 2-day post-confluent 3T3-L1 preadipocytes (day 0) were treated with the indicated concentrations of UVA irradiation and then stimulated with differentiation medium for 3 days. The medium was then replaced with maintenance media every 3 days until the end of the experiment on day 9. The assays were performed on fully differentiated adipocytes (day 9). Intracellular lipids were stained with Oil Red O. The results were confirmed by three independent experiments, which were each conducted in duplicate. E, general viability of cultured cells was determined by the reduction of WST-8 to a highly water-soluble formazan dye. The results were verified by four repetitions of the experiments. F, DNA-damaging effects of UVA irradiation were determined using a DNA Damage ELISA kit. The results were verified by four repetitions of the experiments. G, apoptotic effects of UVA irradiation were determined by Hoechst 33332 staining. The results were verified by four repetitions of the experiments. DIF/MDI, differentiation media.

Article Snippet: Immunoblotting Two-day post-confluent hAMSCs were irradiated with the indicated doses of UVA and then stimulated with STEM PRO® adipocyte differentiation media (Invitrogen) for 3 days.

Techniques: Irradiation, Staining, Activity Assay, Cell Culture, Enzyme-linked Immunosorbent Assay

Journal: The Journal of Biological Chemistry

Article Title: Ultraviolet A Regulates Adipogenic Differentiation of Human Adipose Tissue-derived Mesenchymal Stem Cells via Up-regulation of Kruppel-like Factor 2

doi: 10.1074/jbc.M110.135830

Figure Lengend Snippet: Effects of UVA irradiation on the expression of the adipogenic transcription factors. hAMSCs (day 0) were treated with the indicated doses of UVA irradiation and then stimulated with STEM PRO® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO® adipocyte differentiation media every 3 days until the end of the experiment on day 14. A and D--G, at 1 day or 14 days after the induction of differentiation, the total RNA was isolated, and the mRNA levels of the PPAR γ (A), C/EBP α (D), C/EBP β and C/EBP δ (E), aP2 and LPL (F), and CD36 and LXR α and (G) genes were measured by real time quantitative RT-PCR. The results are expressed relative to untreated cells after normalization against the 18 S rRNA. Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. B, at day 14 after the induction of differentiation, the cell lysates were analyzed by Western blot analysis using the indicated antibodies. The results were verified by three repetitions of the experiments, each of which was conducted in duplicate. C, hAMSCs were cotransfected with luciferase constructs that contained PPRE×3-Luc reporter and PPAR γ cDNA plasmid. After 16 h, the transfected cells were incubated for 20 h with the indicated doses of UVA irradiation. The cells were then harvested and lysed. Luciferase activity is expressed as the ratio of PPRE×3 promoter-dependent firefly luciferase activity divided by control thymidine kinase Renilla luciferase activity (relative luciferase units). Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The experiments were repeated eight times, with each experiment being conducted in triplicate. DIF, differentiation media.

Article Snippet: Immunoblotting Two-day post-confluent hAMSCs were irradiated with the indicated doses of UVA and then stimulated with STEM PRO® adipocyte differentiation media (Invitrogen) for 3 days.

Techniques: Irradiation, Expressing, Isolation, Quantitative RT-PCR, Western Blot, Luciferase, Construct, Plasmid Preparation, Transfection, Incubation, Activity Assay

Journal: The Journal of Biological Chemistry

Article Title: Ultraviolet A Regulates Adipogenic Differentiation of Human Adipose Tissue-derived Mesenchymal Stem Cells via Up-regulation of Kruppel-like Factor 2

doi: 10.1074/jbc.M110.135830

Figure Lengend Snippet: Effects of UVA irradiation on the production of anti-adipogenic cytokines. hAMSCs (day 0) were treated with the indicated doses of UVA irradiation and then stimulated with STEM PRO® adipocyte differentiation media for 3 days. The medium was then replaced with STEM PRO® adipocyte differentiation media every 3 days until the end of the experiment on day 14. At day 14 after the induction of differentiation, the supernatants were harvested for TNF-α (A), IL-1β (B), VEGF (C), TGF-β1 (D), and MIF (E and G) measurements. Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The results were verified by three repetitions of the experiments, each of which was conducted in duplicate. F, at 14 days after the induction of differentiation, the total RNA was isolated, and the mRNA levels of the PPAR γ gene were measured by real time quantitative RT-PCR. The results are expressed relative to untreated cells after normalization against 18 S rRNA. Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The results were verified by four repetitions of the experiments, each of which was conducted in triplicate. DMI, differentiation media; PD, PD98059 (a p42/44 MAPK inhibitor); SB, SB203580 (a p38 MAPK inhibitor); SP, SP600125 (JNK inhibitor); PDTC, pyrrolidine dithiocarbamate.

Article Snippet: Immunoblotting Two-day post-confluent hAMSCs were irradiated with the indicated doses of UVA and then stimulated with STEM PRO® adipocyte differentiation media (Invitrogen) for 3 days.

Techniques: Irradiation, Isolation, Quantitative RT-PCR

Journal: The Journal of Biological Chemistry

Article Title: Ultraviolet A Regulates Adipogenic Differentiation of Human Adipose Tissue-derived Mesenchymal Stem Cells via Up-regulation of Kruppel-like Factor 2

doi: 10.1074/jbc.M110.135830

Figure Lengend Snippet: UVA irradiation has no effects on osteoblast differentiation in hAMSCs. Two-day post-confluent hAMSCs (day 0) were treated with the indicated doses of UVA irradiation and then stimulated with STEM PRO® osteogenesis differentiation media for 3 days. The medium was then replaced with STEM PRO® osteogenesis differentiation media every 3 days until the end of the experiment at day 14. A, alkaline phosphatase staining. Osteogenesis was confirmed with alkaline phosphatase staining. The results were confirmed by three independent experiments, which were each conducted in duplicate. B, alkaline phosphatase activity was measured in cell lysates for 14 days, using a colorimetric assay, and then normalized to the level of total cellular protein. Data are expressed as the means ± S.D. *, p < 0.05 versus controls. The results were confirmed by three independent experiments, which were each conducted in duplicate. ODIF, osteogenic differentiation media.

Article Snippet: Immunoblotting Two-day post-confluent hAMSCs were irradiated with the indicated doses of UVA and then stimulated with STEM PRO® adipocyte differentiation media (Invitrogen) for 3 days.

Techniques: Irradiation, Staining, Activity Assay, Colorimetric Assay

Journal: International Journal of Molecular Sciences

Article Title: Cellular and Biochemical Characterization of Mesenchymal Stem Cells from Killian Nasal Polyp

doi: 10.3390/ijms232113214

Figure Lengend Snippet: Adipocyte differentiation of KNP-MSCs and HNT-MSCs. ( A ) KNP-MSCs and HNT-MSCs were cultured with a standardized adipogenesis differentiation medium and, after 20 days, stained with Oil Red O to detect lipid droplet amount under 40× magnification of phase contrast microscopy. Scale bar: 100 μm. ( B , C ) ImageJ Fiji software analyses were performed to quantify the percentage of red stained area and the mean of red intensity after 20 days of adipogenic differentiation. ( D , E ) Triglyceride-Glo™Assay was performed to assess triacylglycerols and glycerol levels after 20 days of adipogenic differentiation. ( F – I ) The adipogenic differentiation-related gene expression of PPARγ2, FABP4, Adipo-Q, and LPL were evaluated by RT-qPCR after 10 and 20 days in adipogenic differentiation medium, using the housekeeping gene GAPDH for normalization. Data are displayed as means ± SD from three different experiments (** p < 0.001; *** p < 0.0001).

Article Snippet: To induce adipogenic differentiation, 1 × 10 4 cells/cm2 were seeded in 12-well plates and incubated for 20 days with STEM PRO ® Adipogenesis differentiation medium (cat. No. A1007001 Life Technologies).

Techniques: Cell Culture, Staining, Microscopy, Software, Glo Assay, Expressing, Quantitative RT-PCR

Journal: Frontiers in Veterinary Science

Article Title: Canine adipose tissue-derived MSCs engineered with mRNA to overexpress TSG-6 and enhance the anti-inflammatory effects in canine macrophages

doi: 10.3389/fvets.2023.1134185

Figure Lengend Snippet: Characterization of cAT-MSCs isolated from canine adipose tissue. (A–C) Trilineage potential of cAT-MSCs. Differentiation into adipocyte, osteocyte, and chondrocyte was confirmed via Oil Red O, Alizarin Red, and Alcian Blue staining, respectively. (D–G) Flow cytometry analysis of cell surface markers. cAT-MSCs expressed CD29 and CD90 and lacked CD34 and CD45. (Blue: stained marker, red: unstained negative control).

Article Snippet: To assess their differentiation capacities, cAT-MSCs were cultured using three differentiation media (StemPro Adipogenesis Differentiation, Stem Pro Osteogenesis Differentiation, and StemPro Chondrogenesis Differentiation kits; Gibco).

Techniques: Isolation, Staining, Flow Cytometry, Marker, Negative Control

Journal: Frontiers in Veterinary Science

Article Title: Canine adipose tissue-derived MSCs engineered with mRNA to overexpress TSG-6 and enhance the anti-inflammatory effects in canine macrophages

doi: 10.3389/fvets.2023.1134185

Figure Lengend Snippet: cAT-MSCs expressing GFP with three different UTRs. (A) Sequences of HBA1, HBA2 , and HBB UTR regions. (B) Schematic structure of each template DNA, consisting of a T7 promoter, a 5′-UTR, a GFP coding region, and a 3′-UTR. (C) The highest fluorescence level was observed with HBA1 24 h post-transfection. HBA2 and HBB reached peak fluorescence levels at 12 and 72 h post-transfection, both at lower levels than those of HBA1 . Fluorescence was observed up to 96 h post-transfection. Scale bar, 400 μm ( HBA1 : GFP mRNA with UTR of canine hemoglobin subunit alpha-like 1; HBA2 : GFP mRNA with UTR of canine hemoglobin subunit alpha-like 2; HBB : GFP mRNA with UTR of canine hemoglobin subunit beta-like).

Article Snippet: To assess their differentiation capacities, cAT-MSCs were cultured using three differentiation media (StemPro Adipogenesis Differentiation, Stem Pro Osteogenesis Differentiation, and StemPro Chondrogenesis Differentiation kits; Gibco).

Techniques: Expressing, Fluorescence, Transfection

Journal: Frontiers in Veterinary Science

Article Title: Canine adipose tissue-derived MSCs engineered with mRNA to overexpress TSG-6 and enhance the anti-inflammatory effects in canine macrophages

doi: 10.3389/fvets.2023.1134185

Figure Lengend Snippet: TSG-6 mRNA transfection increased expression of TSG-6 in cAT-MSCs. (A) TSG-6 mRNA level was increased in cAT-MSCs transfected with TSG-6 mRNA (MSC TSG-6 ). (B) TSG-6 protein level was increased in MSCs TSG-6 . (C) Secretion of TSG-6 was also increased in MSCs TSG-6 . Results are shown as the mean ± standard deviation. ** P < 0.01, **** P < 0.0001.

Article Snippet: To assess their differentiation capacities, cAT-MSCs were cultured using three differentiation media (StemPro Adipogenesis Differentiation, Stem Pro Osteogenesis Differentiation, and StemPro Chondrogenesis Differentiation kits; Gibco).

Techniques: Transfection, Expressing, Standard Deviation