中文  |  EN
Reagents
hPSC-Dopaminergic Neuron Differentiation Kit
hPSC-Dopaminergic Neuron Differentiation Kit
hPSC-Dopaminergic Neuron Differentiation Kit
hPSC-Dopaminergic Neuron Differentiation Kit

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.

hPSC-mDAP Differentiation Kit can differentiate hPSC into high-purity mDAP (midbrain dopaminergic progenitors) (>90% Lmx1a+/ Foxa2+/ En1+). hPSC-mDAN Maturation Medium can differentiate mDAP into mature mDAN (TH+/ Nurr1+), has mDAN-specific electrophysiological activity and the function of releasing dopamine. The differentiated human mDAP and mDAN can be used in scientific research of neurodegenerative diseases, drug screening, and cell transplantation of Parkinson's disease models.

Contents Amount Cat. No. Storage

Nuwacell® hPSC- Dopaminergic Neuron Differentiation Kit

     

Nuwacell® hPSC-mDAP Differentiation Kit*

1 Kit    RP01017

Basal medium

  2℃~8℃

Supplement 

-20℃~-80℃

Nuwacell® hPSC-mDAP Maintenance Medium

100 mL RP01017-H

Nuwacell® hPSC-mDAN Maturation Medium

50 mL RP01017-l

* 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.

 

1、Process

 

 

Schematic of the hPSC-derived mDAN differentiation using a hPSC-Dopaminergic Neuron Differentiation Kit.

2、Cell morphology during differentiation

 

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.

3、Cell specific markers of mDAP

 

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.

4、Cell specific markers expression and functional analysis of mDAN

 

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.

Copyright © 2016-2022 Nuwacell Biotechnologies Co., Ltd. All rights reserved. ICP:16011195
Public security record No 34019202001635 Clause