Nuwacell® hPSC-Dopaminergic Neuron Differentiation Kit（RP01017）, originally developed by Nuwacell Biotechnologies Co., Ltd. is suitable for the highly effective differentiation of human pluripotent stem cells (hPSC) into mature mDAN (midbrain dopaminergic neurons). The kit includes hPSC-mDAP Differentiation Kit, hPSC-mDAP Maintenance Medium, and hPSC-mDAN Maturation Medium, which can be flexibly selected according to specific needs.
Nuwacell® hPSC- Dopaminergic Neuron Differentiation Kit
Nuwacell® hPSC-mDAP Differentiation Kit*
Nuwacell® hPSC-mDAP Maintenance Medium
Nuwacell® hPSC-mDAN Maturation Medium
* Human mDAP (midbrain dopaminergic progenitors) can be obtained at approximately 1×107 cells on day 11 from a hPSC-Dopaminergic Neuron Differentiation Kit.
* The complete hPSC-Dopaminergic Neuron Differentiation Medium can be stored at 2-8°C for up to 2 weeks.
Schematic of the hPSC-derived mDAN differentiation using a hPSC-Dopaminergic Neuron Differentiation Kit.
The morphology of hPSC-derived DA cultured at day 0 (hiPSC), day 11 (mDAP), day 12 (subculture of mDAP), and day 45 (mDAN) during differentiation. Scale bar, 200 μm.
Immunofluorescence staining of LMX1A, FOXA2, and EN1 for differentiated mDAP using a hPSC-mDAP Differentiation Kit. Scale bar, 200 μm.
(A)Quantitative PCR of NKX2.1 (a diencephalon marker), DBX1 (a midbrain marker), LMX1a (a midbrain marker), FOXA2 (a midbrain marker), EN1 (a midbrain marker), OTX2 (a midbrain marker), and GBX2 (a hindbrain marker). (B) Representative flow cytometry analysis of FOXA2/En1 and FOXA2/LMX1a.
Immunofluorescence staining of TH and Nurr1 for differentiated mDAN using a hPSC-Dopaminergic Neuron Differentiation Kit. Scale bar, 200 μm.
Radioimmunoassay of dopamine release level of differentiated mDAN (at day 45), which is higher than that from hPSC control (i.e., >2000 pg/mL versus）
Excitability analysis of hPSC-mDAN (at day 42) by electrophysiology
A representative image of electrophysiological recording on individual mDAN. Scale bar, 20 μm. Representative traces of spontaneous action potentials at 2-10 Hz in the current-clamp mode. A slowly increasing inward current (H current) were reliably elicited from a hyperpolarized voltage in the voltage-clamp mode.