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Stimulation of L-type calcium channels increases tyrosine hydroxylase and dopamine in ventral midbrain cells induced from somatic cells

Stem Cells Transl Med. 2020;

Jefri M, Bell S, Peng H, Hettige N, Maussion G, Soubannier V, Wu H, Silveira H, Theroux JF, Moquin L, Zhang X, Aouabed Z, Krishnan J, O'Leary LA, Antonyan L, Zhang Y, McCarty V, Mechawar N, Gratton A, Schuppert A, Durcan TM, Fon EA, Ernst C, .

Products/Services Used	Details	Operation
Recombinant Proteins	After 1 week, when cells reached maximum confluence, mNIM1 was switched to mNIM2 (DMEM/F12 supplemented with L-glutamine [2 mM], N2, B27 supplement, bovine serum albumin [1 mg/mL, ThermoFisher], NEAA [10 mM; ThermoFisher], SHH C24II [200 ng/mL; GenScript], CHIR-99021 [STEMCELL Technologies], and laminin [1 μg/mL, Sigma-Aldrich]).... When 15%-25% confluence was reached, iPSC culture media was replaced with neural induction (NIM1) medium 1 (DMEM/F12 supplemented with L-glutamine [2 mM], N2, B27 supplement, bovine serum albumin [1 mg/mL, Ther- moFisher], NEAA [10 mM; ThermoFisher], SB431542 [10 μM; STEMCELL Technologies], noggin [200 ng/mL, GenScript], and lami- nin [1 μg/mL, Sigma-Aldrich]).	Get A Quote

摘要

Making high-quality dopamine (DA)-producing cells for basic biological or small molecule screening studies is critical for the development of novel therapeutics for disorders of the ventral midbrain. Currently, many ventral midbrain assays have low signal to noise ratio due to low levels of cellular DA and the rate-limiting enzyme of DA synthesis, tyrosine hydroxylase (TH), hampering discovery efforts. Using intensively characterized ventral midbrain cells derived from human skin which demonstrate calcium pacemaking activity and classical electrophysiological properties, we show that an L-type calcium agonist can significantly increase TH protein levels and DA content and release. Live calcium imaging suggests ... More