Human induced pluripotent stem cells (h-iPSCs) were generated and cultured as previously described [49 (link)]. In brief, h-iPSCs were generated through non-integrating episomal overexpression of reprogramming factors OCT4, SOX2, KLF4, L-MYC, and LIN28 into human MSCs via electroporation. H-iPSCs were cultured on Matrigel (Corning, Cat No. 354277) coated plate in mTeSR1 medium (STEMCELL, Cat No. 85850) and passaged when reaching 80% confluency. Cardiac differentiation was induced using STEMdiff™ cardiomyocyte differentiation kit (STEMCELL, Cat No. 05010) according to the manufacturer’s protocol. Following 8 days of differentiation, metabolic selection was performed with lactate medium composed of RPMI 1640 medium (no glucose, ThermoFisher, Cat No. 11879020) supplemented with 1% sodium DL-lactate solution (60%, Sigma, Cat No. L4263) and B27 (minus insulin, ThermoFisher, Cat No. A1895601). After 2 days of lactate selection, purified iCMs were maintained in STEMdiff™ cardiomyocyte maintenance medium.
Dl lactate solution
Manufactured by Merck Group
DL-lactate solution is a sterile, aqueous solution containing a mixture of D-lactate and L-lactate. The solution is intended for use as a laboratory reagent.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using dl lactate solution
1
Generating Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes
2
Metabolic Inhibitors in Embryonic Assay
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The pharmacological inhibitors used in this study include Hexokinase inhibitor 2DG (2-Deoxy-Glucose, Sigma D8375), PFKFB3 inhibitor 3PO (Sigma SML1343), Pyruvate Dehydrogenase Kinase inhibitor DCA (dichloroacetate, Sigma 347795), Lactate Dehydrogenase inhibitor Galloflavin (Sigma SML0776), MCT1/2/4 inhibitors CHC (á-cyano-4-hydroxycinnamic acid, Sigma C2020) and UK-5099 (Sigma PZ0160), MEK inhibitor U1026 (Sigma U120) and PI3K inhibitor LY294002 (Sigma L9908). All inhibitors were dissolved in a 10 mm DMSO stock solution and diluted into the final indicated concentrations in fish water. Lactate treatments were performed with a 60% stock sodium DL-lactate solution (Sigma L1375) diluted 1 to 1000 in fish water to a final concentration of 6.7 mM, and buffered with 10 mm MES at pH 6.2. Note, lactate treatment has little effect on embryonic development at higher pH (not shown). Treatments were carried in a 24-well plate with a maximum of 15 embryos per well in a volume of 500 µl each.
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