Clinical and histologic scoring of mice was monitored as previously reported [19 (link), 27 (link)]. The survival, body weight, and appearance of mice in each group were assessed before sacrifice at the indicated time point. The clinical score of GVHD was evaluated using a clinical scoring system as shown in Additional file 4 : Figure S4a. For histologic scoring of mice, the sections of the liver, lung, and skin were stained with hematoxylin-eosin (H&E) staining and observed under a Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan) as we described before [26 (link)]. Each pathological index of the lung, liver, or skin was graded from 0 to 3, and the total GVHD histologic index was the sum of the parameters listed in Additional file 5 : Table S1.
Elipseti u microscope
Manufactured by Nikon
Sourced in Japan
The ElipseTi-U microscope is a high-performance laboratory instrument designed for advanced microscopic analysis. It features a sturdy construction, precision optics, and a range of customizable features to suit diverse research and testing requirements. The ElipseTi-U microscope provides reliable and accurate imaging capabilities to support various scientific and industrial applications.
Automatically generated - may contain errors
Lab products found in correlation
Von kossa staining, by Abcam (1 mentions)
H e staining, by Merck Group (1 mentions)
Mesencult adipogenic differentiation kit, by STEMCELL (1 mentions)
Mesencult acf chondrogenic differentiation kit, by STEMCELL (1 mentions)
Mesencult osteogenic differentiation kit, by STEMCELL (1 mentions)
Alcian blue, by Abcam (1 mentions)
7 protocols using elipseti u microscope
1
Histological Scoring of GVHD in Mice
2
Osteogenic Differentiation of ASCs
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Osteogenic differentiation was conducted as we previously described with several modification (10 (link)). In details, ND-ASCs and T2DM-ASCs were seeded at a density of 2×104 cells/cm2 at 37℃, 5% CO2. When cells reached 80% confluent, the supernatant was discarded and changed into osteogenic differentiation medium containing 100 nM dexamethasone, 50 μM L-ascorbic acid 2-phosphate, and 10 mM β-glycerophosphate in Serum-free human MSC medium (Yocon). The medium was subsequently changed twice a week. After 28 days, osteogenic differentiation was assessed by von Kossa staining (Abcam). The phase contrast image was photographed with Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan).
3
Cell Migration Scratch Assay
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ND-ASCs or T2DM-ASCs were seeded in 6-well plate, when the ND-ASCs or T2DM-ASCs reached 80% confluent, the indicated cell layers were scratched with 200 μl tips and captured with a Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan) at the indicated time points (0 h, 12 h, 24 h). Images of the scratch area were captured and analyzed by the ImageJ software.
4
Histochemical Tissue Staining Protocol
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H&E staining (Sigma-Aldrich) and histochemical staining (Abcam, Cambridge, UK) were performed according to the manufacturers’ instructions. Samples were photographed using a Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan).
5
Adipogenic Differentiation of Stem Cells
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Adipogenic differentiation was conducted as we recently reported with several modification (10 (link)). In details, ND-ASCs and T2DM-ASCs were seeded at a density of 2×104 cells/cm2 at 37℃, 5% CO2. When cells reached 80% confluent, the supernatant was discarded and changed into adipogenic differentiation medium containing 1 μM dexamethasone, 0.1 mM indomethacin, 0.5 mM 3-isobutyl-1-methylxanthine, and 10 mg/L insulin in Serum-free human MSC medium (Yocon). The medium was subsequently changed twice a week. After 21 days, adipogenic differentiation was assessed by Oil Red O staining of the fat droplets. The phase contrast image was photographed with Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan).
6
Tri-lineage Differentiation of hUC-MSCs
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hUC-MSCs at various passages (P3, P6, P15) were seeded at a density of 2 × 104/cm2 in MSC culture medium. When cells reached 80% fusion, the medium was changed into adipogenic (MesenCult Adipogenic Differentiation Kit, Stem Cell Technologies), osteogenic (MesenCult Osteogenic Differentiation Kit, Stem Cell Technologies), or chondrogenic (MesenCult-ACF Chondrogenic Differentiation Kit, Stem Cell Technologies) differentiation medium. The differentiation medium was changed every 3 days as we described previously [6 (link), 20 (link)]. Twenty-one days later, the hUC-MSC-derived cells were stained by Oil Red S, Alizarin Red, or Alcian Blue staining and photographed with Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan). The primer sequences for tri-lineage differentiation are available in Additional file 7 : Table S1.
7
Chondrogenic Differentiation of ASCs
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Chondrogenic differentiation was performed as we recently reported (10 (link)). Briefly, ND-ASCs and T2DM-ASCs were seeded at a density of 2×104 cells/cm2 at 37℃, 5% CO2. When cells reached 80% confluent, the supernatant was discarded and changed into chondrogenic differentiation medium (StemCells). The medium was subsequently changed twice a week. After 28 days, chondrogenic differentiation was assessed by Alcian Blue staining (Abcam). The phase contrast image was photographed with Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan).
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