Recombinant human insulin (29 IU/mg) was purchased from Dongbao Enterprise Group Co., Ltd. (Tonghua, China). Lecithin (72% phosphatidylcholine/18% phosphatidylethanolamine, Lipoid S75) was obtained from Shanghai Tywei Pharmaceutical Co., Ltd. (Shanghai, China). Tween-20 was purchased from China National Medicines Co., Ltd. (Beijing, China). Bile acids sodium salt was obtained from Anhui Chem-Bright bioengineering Co., Ltd. (Anhui, China). Gelatin was purchased from Shandong Hengxin Biotechnology Co., Ltd. (Shandong, China). FITC-insulin was obtained from Melone Pharmaceutical Co., Ltd. (Liaoning, China). DAPI and TRITC phalloidin were purchased from Beyotime Biotechnology Co., Ltd. (Shanghai, China). Other chemicals and solvents were of analytical or chromatography grade.
Tritc phalloidin
Manufactured by Beyotime
Sourced in China
TRITC phalloidin is a fluorescent dye used to label F-actin, a type of actin filament found in the cytoskeleton of cells. It binds to F-actin with high affinity, allowing for the visualization and analysis of actin structures within cells.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Beyotime (4 mentions)
Glass bottom dish, by Corning (1 mentions)
Sp8x confocal laser scanning microscope, by Leica (1 mentions)
4 6 diamidino 2 phenylindole dihydrochloride dapi, by Beyotime (1 mentions)
Triton x 100, by Keygen Biotech (1 mentions)
Propidium iodide, by Beyotime (1 mentions)
Cell counting kit 8 cck 8, by Beyotime (1 mentions)
Calcein am, by Beyotime (1 mentions)
Tributyl phosphate, by Merck Group (1 mentions)
3 3 cholamidopropyl dimethylammonio 1 propane sulfonate, by Merck Group (1 mentions)
Triton x 100 solution, by Merck Group (1 mentions)
Benzonase nuclease, by Merck Group (1 mentions)
Amidosulfobetaine 14 asb 14, by Merck Group (1 mentions)
Cell counting kit 8 cck 8, by Biosharp (1 mentions)
Fv3000, by Olympus (1 mentions)
Cell counting kit 8 cck 8, by BestBio (1 mentions)
Live dead cell staining kits, by BestBio (1 mentions)
7 protocols using tritc phalloidin
1
Insulin-loaded Phospholipid Nanocarriers
2
Cellular Uptake and Imaging of Nanoliposomes
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TR146 cells were seeded in a glass-bottom dish (Corning) at a density of 5 × 104 cells/dish and incubated at 37 °C under 5% CO2 for 24 h to allow the cells to adhere. The culture medium was discarded and replaced with fresh, serum-free medium containing serially diluted nanoliposomes (100 μg/mL, final concentration). The cells were incubated at 37 °C under 5% CO2 for 2 h, washed three times with pre-chilled PBS at 4 °C, and fixed with 4% paraformaldehyde in PBS (w/v) for 20 min at room temperature (RT). The cells were rinsed three times with PBS and permeabilized with 0.5% Triton X-100 for 5 min at RT. The cells were then washed three times with PBS, stained with 500 μL TRITC phalloidin (Beyotime Biotechnology), and incubated for 50 min at RT. After washing three times with PBS, the cells were counterstained with 500 μL DAPI and stored at 4 °C. Fluorescence was observed under an SP8X confocal laser scanning microscope (Leica, Wetzlar, Germany). DAPI (green fluorescence) was measured at excitation and emission wavelengths of 364 and 454 nm. TRITC (red fluorescence) was measured at excitation and emission wavelengths of 545 and 570 nm. FITC-labeled insulin (green fluorescence) was measured at excitation and emission wavelengths of 490 and 525 nm.
3
Adhesion of hDPSCs to Material
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This study evaluated the adhesion of hDPSCs to the material. As mentioned above, a cylinder with a diameter of 10 mm and a thickness of 1 mm was prepared by covering the cell climbing sheet with material on a super-clean table and placed in a cell incubator with humidity greater than 95% and temperature of 37 ℃. After 1 week, the cells were placed in 24-well plates with 4 compound pores in each group. Inoculate hDPSCs on the surface of the 24-well plates (1×104 cells per well). After 24h and 72h, the cell scaffold complexes were fixed with 4% paraformaldehyde for 30 minutes and permeated with 0.5% Triton X-100 (KeyGEN BioTECH) for 5 minutes at room temperature. TRITC Phalloidin and 4,6-diamidino-2-phenylindole dihydrochloride (DAPI, Beyotime) were used to label actin and nucleus. Images were acquired using an inverted phase-contrast fluorescence microscope (IX-71,Olympus) to observe the attachment of hDPSCs on the scaffold. At the same time, SEM was used to observe the changes of the cells inoculated for 72 hours on the cell slide, Images were captured to visualise the cells adhering to the scaffold.
4
Gelatin-Based Hydrogel for Stem Cell Culture
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Gelatin (type A, from porcine skin), methacrylic anhydride (MA), and sodium periodate (NaIO4) were obtained from Aladdin Industrial Corporation (Shanghai, China). Dextran (Mw = 70 kDa) was purchased from J&K Scientific co., Ltd (Beijing, China). Lysozyme was purchased from commercially Shanghai Yuanye Bio-Technology Co., Ltd. (Shanghai, China). Human umbilical cord mesenchymal stem cells (UCMSCs, AC340316) was purchased from American Type Culture Collection (Manassas, VA). Live/Dead cell staining kits, Cell Counting Kit-8 (CCK-8), Calcein-AM, propidium iodide, 4,6-diamidino-2-phenylindole (DAPI) and TRITC Phalloidin were purchased from Biyuntian Biotechnology CO. LTD. (Shanghai, China). All of the chemical reagents were of analytical grade and used without any further purification.
5
Biocompatible PVA Scaffold Characterization
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Powder PVA (MW: 7.2–8.14 × 104 g/mol) was purchased from Yingjia Industrial Development Co., Shanghai, China. Cell counting kit-8 (CCK-8) was obtained from Biosharp, Hefei, China. Phosphate buffered saline (PBS), Penicillin/streptavidin (PS), and Trypsin were purchased from HyClone, South Logan, UT, USA. 4’,6-diamidino-2-phenylindole (DAPI) fluorescent staining solution, Calcein-AM/PI Live Dead Cell Assay Kit, TRITC-Phalloidin and Tris(hydroxymethyl)aminomethane hydrochloride (TRIS-HCI) Buffer were purchased from Biyuntian Biotechnology Co., Shanghai, China. TritonX-100 solution, 3-[(3-Cholamidopropyl)-dimethyl-ammonio]-1-propane sulfonate (CHAPS), Tributyl phosphate, N-Decyl-N, N-dimethyl-3-ammonio-1-propanesulfonate (SB3-10), Amidosulfobetaine-14 (ASB-14) and Benzonase nuclease were obtained from Sigma-Aldrich, St. Louis, MO, USA.
6
Actin and Nuclei Staining Protocol
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TRITC phalloidin (Biyuntian Biotechnology Co., Ltd., China) and 4, 6‐diamidino‐2‐phenylindole, known as DAPI (Biyuntian Biotechnology Company, China), were used as a fluorescent dye to stain filamentous actin (F‐actin) and nuclei. In brief, the cells were rinsed with PBS (1 min for three times) followed by immersed in 4% paraformaldehyde for 30 min to fixed cells. Then, the cells were immersed in 0.5% (v:v) Triton X‐100 to increase the membrane permeability for 10 min. After rinsed for three times, the cells were stained with TRITC phalloidin for 30 min before the nuclei was stained with DAPI for 10 min in dark at room temperature. Lastly, the cell morphology of hydrogels was observed by fluorescence microscope (Olympus FV3000).
7
Gelatin-Collagen Hydrogel for Stem Cell Cultivation
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Gelatin (Gel, BioReagent,
from cold water fish skin) was purchased from Sigma-Aldrich Co., Ltd.
(Darmstadt, Germany). Collagen (Col, type I collagen, MW = 100–200
kDa) was obtained from Haishen Biotechnology Co., Ltd. (Fuzhou, China).
Methacrylic anhydride (MA) was purchased from Maclean Biochemical
Technology Co., Ltd. (Shanghai, China). A photoinitiator (lithium
phenyl-2,4,6-trimethylbenzoylphosphinate, LAP) was purchased from
Shanghai Yingchang Biotechnology Co., Ltd. (Shanghai, China). Cell
Counting Kit-8 (CCK-8) and Live/Dead cell staining kits were purchased
from BestBio Co., Ltd. (Shanghai, China). TRITC phalloidin and 4,6-diamidino-2-phenylindole
(DAPI) were obtained from Biyuntian Biotechnology Co., Ltd. (Shanghai,
China). Human amnion mesenchymal stem cells (hAMSCs) derived from
a human amnion were purchased from Fuyuan Biotechnology Co., Ltd.
(Shanghai, China). Unless otherwise stated, all chemicals were of
analytical reagent grade.
from cold water fish skin) was purchased from Sigma-Aldrich Co., Ltd.
(Darmstadt, Germany). Collagen (Col, type I collagen, MW = 100–200
kDa) was obtained from Haishen Biotechnology Co., Ltd. (Fuzhou, China).
Methacrylic anhydride (MA) was purchased from Maclean Biochemical
Technology Co., Ltd. (Shanghai, China). A photoinitiator (lithium
phenyl-2,4,6-trimethylbenzoylphosphinate, LAP) was purchased from
Shanghai Yingchang Biotechnology Co., Ltd. (Shanghai, China). Cell
Counting Kit-8 (CCK-8) and Live/Dead cell staining kits were purchased
from BestBio Co., Ltd. (Shanghai, China). TRITC phalloidin and 4,6-diamidino-2-phenylindole
(DAPI) were obtained from Biyuntian Biotechnology Co., Ltd. (Shanghai,
China). Human amnion mesenchymal stem cells (hAMSCs) derived from
a human amnion were purchased from Fuyuan Biotechnology Co., Ltd.
(Shanghai, China). Unless otherwise stated, all chemicals were of
analytical reagent grade.
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