Carnitine hydrochloride

Manufactured by Merck Group

Sourced in United Kingdom

Carnitine hydrochloride is a water-soluble, organic compound that is used in various laboratory applications. It acts as a cofactor in the transport of long-chain fatty acids into the mitochondria for beta-oxidation.

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Lab products found in correlation

3 protocols using carnitine hydrochloride

Fatty Acid Oxidation Assay in BMDMs

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The ability of the cells to oxidize exogenous fatty acid substrates was measured using the Agilent Seahorse XF Cell Mito Stress Test kit according to manufacturer’s recommendations and the data analyzed using the Agilent Seahorse XF Wave software tool as described in the previous section. BMDMs (4x10⁵ cells) were seeded in Agilent Seahorse XF96 cell culture microplate and allowed to adhere in complete culture medium. The growth medium was then replaced and incubated for 24 hours with substrate limited DMEM medium containing reduced concentrations of glucose (0.5mM), glutamax (1mM) and fetal bovine serum (1%) to deplete endogenous substrates within the cells. Carnitine hydrochloride (0.5mM) (Sigma, St. Louis, MO) was added fresh during the media change. The media was then replaced with FAO assay medium containing 111mM NaCl, 4.7mM KCl, 1.25mM CaCl₂, 2mM MgSO4, 1.2mM NaH₂PO₄ supplemented with 2.5mM glucose, 0.5mM Carnitine and 5mM HEPES and incubated for 30-45 minutes. XF Palmitate BSA or BSA control was added to the medium just prior to the start of the XF assay. The OCR (pmol of O₂/min) was measured after sequential addition of electron transport chain modulators Oligomycin, FCCP and Rotenone/Antimycin A.

Wessendarp M., Watanabe-Chailland M., Liu S., Stankiewicz T., Ma Y., Kasam R.K., Shima K., Chalk C., Carey B., Rosendale L.R., Filippi M.D, & Arumugam P. (2021). Role of GM-CSF in regulating metabolism and mitochondrial functions critical to macrophage proliferation. Mitochondrion, 62, 85-101.

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Co-culture of hiPSC-CM and adult rat cardiomyocytes

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The hiPSC-CM were co-cultured with left-ventricle cardiomyocytes, freshly isolated following the method outlined in^{23 (link)} from male rats, anesthetized with 5% isoflurane prior to sacrifice. The cardiomyocytes were suspended in low Ca²⁺ enzyme solution, containing (in mM) NaCl (120), KCl (5), MgSO₄ (5), Na pyruvate (5), glucose (5), taurine (20), HEPES (10), CaCl₂ (0.2), pH 7.4 adjusted with 1 M NaOH. The adult-CM were re-suspended at 1:10 dilution in M199 + culture medium containing (per 500 ml M199): 5 mL P/S, 1 g Bovine Serum Albumin (BSA, Sigma-Aldrich), 0.33 g creatinine (Sigma-Aldrich), 0.33 g taurine (Sigma-Aldrich), 0.0088 g l-ascorbic acid (Scientific Laboratory Supplies), 0.161 g carnitine hydrochloride (Sigma-Aldrich). To make the co-culture, the RPMI cell media was aspirated almost completely from the hiPSC-CM culture and quickly replaced with the same amount of M199 + culture medium. Next, suspended isolated adult-CM were added dropwise on top of the stem cell culture to achieve a scattered adult-CM distribution (Supplementary Fig. 1a). The cells were co-cultured at 5% CO₂, 37 °C for varying durations (4 h, 1 day, 2 days) up until the point of imaging, with daily media changes.

Dvinskikh L., Sparks H., Brito L., MacLeod K.T., Harding S.E, & Dunsby C. (2023). Remote-refocusing light-sheet fluorescence microscopy enables 3D imaging of electromechanical coupling of hiPSC-derived and adult cardiomyocytes in co-culture. Scientific Reports, 13, 3342.

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Cardiomyocyte contractility regulation

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M199 medium (Invitrogen UK, 11150), taurine (Biochemica, A1141), creatine monohydrate (Sigma Aldrich, C3630), penicillin/streptomycin (Merck, A2212), carnitine hydrochloride (Sigma Aldrich, C9500) BSA (Sigma Aldrich, A6003), laminin (Sigma Aldrich L2020), isoproterenol hydrochloride (Sigma Aldrich, I6504), ICI118551 (Tocris UK, 0821), CGP 20712A (Tocris UK, 1024), SR 59230A hydrochloride (Tocris UK, 1511), L-NAME hydrochloride (Tocris UK, 0665), Vinpocetine (Sigma Aldrich, V6383), EHNA hydrochloride (Sigma Aldrich, E114), Cilostamide (Tocris UK, 0915), Tadalafil (Santa Cruz USA, sc-208412), IBMX (Santa Cruz sc-201188), self-made rabbit sGCα and β subunit antibodies (specificity tested in KO animals; Friebe et al., 2018 (link)), mouse α-actinin (Sigma Aldrich, A7732), mouse Caveolin-3 (BD Transduction Laboratories, 610421, specificity tested in KO animals; Woodman et al., 2002 (link)), secondary Alexa Fluor antibodies 488 nm, 514 nm, 568 nm and 633 nm (Life Technologies), BSA (Fisher Scientific UK, BPE9704), fluorescence mounting medium (Vectashield Germany, H-1000), MaTek glass-bottom dishes (MaTek USA, P35G-1.5–10 C), TAT-scram and TAT-Cav3 peptides (a gift from Dr. Sarah Calaghan from Leeds, England).

Schobesberger S., Wright P.T., Poulet C., Sanchez Alonso Mardones J.L., Mansfield C., Friebe A., Harding S.E., Balligand J.L., Nikolaev V.O, & Gorelik J. (2020). β3-Adrenoceptor redistribution impairs NO/cGMP/PDE2 signalling in failing cardiomyocytes. eLife, 9, e52221.

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