EDL muscles were removed carefully and digested with 2 mg ml−1 collagenase type 1 (Worthington) in Dulbecco’s Modified Eagle’s Medium (DMEM, Gibco) for 45 min−1 h at 37 °C. Digestion was stopped by carefully transferring EDL muscles to a horse serum-coated Petri dish (60-mm) with DMEM. Myofibers were released by gently flushing muscles with a large bore glass pipette. Released single myofiber was either fixed immediately with 4% PFA or transferred and cultured in a horse serum-coated Petri dish (60-mm) in DMEM supplemented with 20% fetal bovine serum (FBS, Gibco), 4 ng ml−1 basic fibroblast growth factor (bFgf, Peprotech), and 1% penicillin–streptomycin (P/S, Gibco) at 37 °C for indicated days following the protocol by Yue and colleagues46 (link). Dox (RPI) was administered in culture medium at a concentration of 1 µg ml−1 to induce OSKM expression. Recombinant Wnt4 (R&D systems) was administered in culture medium at a concentration of 100 ng ml−1 according to a previous study23 .
Recombinant wnt4
Manufactured by R&D Systems
Recombinant Wnt4 is a protein produced using recombinant DNA technology. It is a member of the Wnt family of secreted signaling proteins, which play crucial roles in various biological processes. The core function of Recombinant Wnt4 is to facilitate the study of Wnt signaling pathways and their underlying mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using recombinant wnt4
1
Muscle Fiber Isolation and Culture
2
Chondrogenic and Osteogenic Differentiation of MSCs
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For chondrogenesis, MSCs were cultured as a pellet in chondrogenic medium (DMEM supplemented with 1 U/mL penicillin, 1 μg/mL streptomycin, 100 μg/mL sodium pyruvate, 100 nM dexamethasone, 50 μg/mL L-ascorbate-2-phosphate, 1X ITS-1 premix, 10 ng/mL TGFβ3). For histology, either 500,000 (large) or 100,000 (mini) MSCs were seeded into a pellet. For gene expression analysis, 150,000 MSCs were seeded into a pellet. Chondrogenic media was replaced every 2 to 3 days for 14 days. For osteogenesis, MSCs were seeded in monolayer and induced to differentiate in osteogenic media as outlined above. Osteogenic media was replaced every 2 to 3 days for 21 days. For WNT4 recovery studies, recombinant WNT4 (R&D Systems) was added at a concentration of 100 ng/mL and was added to the culture from day 4 (chondrogenesis) and day 7 (osteogenesis).
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