STEMdiff™ Midbrain Neuron Differentiation Kit_产品中心

细胞培养基及添加物

STEMdiff™ Midbrain Neuron Differentiation Kit

Differentiation kit for the generation of midbrain-patterned neuronal precursors from human ES and iPS cell-derived neural progenitor cells

概要

技术资料

数据及文献

概要

STEMdiff™ Midbrain Neuron Differentiation Kit (Catalog #100-0038) is used to generate midbrain neuronal precursors from neural progenitor cells (NPCs) derived from human pluripotent stem cells (hPSCs) using STEMdiff™ SMADi Neural Induction Kit (Catalog #08581). The midbrain neuronal precursors are further matured into midbrain neurons using STEMdiff™ Midbrain Maturation Kit (Catalog #100-0041). These media will produce a population of midbrain neurons (≥ 15% TH-positive dopaminergic neurons; ≥ 90% class III βtubulin-positive neurons; < 10% GFAP-positive astrocytes). Cells derived using these products are versatile tools for modeling human neurological development and disease, drug screening, toxicity testing, and cell therapy validation.

技术资料

Document Type	产品名称	Catalog #	Lot #	语言
Product Information Sheet	STEMdiff™ Midbrain Neuron Differentiation Kit	100-0038	All	English
Safety Data Sheet 1	STEMdiff™ Midbrain Neuron Differentiation Kit	100-0038	All	English
Safety Data Sheet 2	STEMdiff™ Midbrain Neuron Differentiation Kit	100-0038	All	English

数据及文献

Data

Experimental Protocol for STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits (Embryoid Body Protocol)

Figure 1. Schematic for the Embryoid Body Protocol

Midbrain-type neural precursors can be generated in 18 - 19 days from hPSC-derived neural progenitor cells (NPCs) after selecting neural rosettes from replated embryoid bodies. For the maturation of precursors to midbrain-type neurons, including dopaminergic neurons, see the PIS.

Experimental Protocol for STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits (Monolayer Protocol)

Figure 2. Schematic for the Monolayer Protocol

Midbrain-type neural precursors can be generated from neural progenitor cell (NPC) monolayers derived from embryonic and induced pluripotent stem cells after three single-cell passages. For the maturation of precursors to midbrain-type neurons, including dopaminergic neurons, see the PIS.

Midbrain-Type Neurons Arise From Neural Progenitor Cells After Culture in STEMdiff™ SMADi Neural Induction Kit and STEMdiff™ Midbrain Culture System

Figure 3. Midbrain-Type Neurons Are Generated After Culture in STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits

NPCs generated from H1 hPSCs in mTeSR™1 using the STEMdiff™ SMADi Neural Induction Kit embryoid body (EB) protocol were differentiated and matured to midbrain-type neurons using the STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits. (A) Midbrain-type neurons were formed after hPSC-derived NPCs were cultured with the STEMdiff™ Midbrain Neuron Differentiation Kit for 12 days and STEMdiff™ Midbrain Neuron Maturation Kit for 14 days. The resulting cultures contain a population of (B) class III β-tubulin-positive neurons (magenta), with (D) more than 15% Tyrosine Hydroxylase-positive cells (green). (C) Nuclei are labeled with DAPI (blue).

Figure 4. Midbrain-Type Neural Precursor Cells Express Characteristic Markers After Culture in STEMdiff™ Midbrain Neuron Differentiation Kit

NPCs generated from STiPS-F016 hPSCs in mTeSR™1 using the STEMdiff™ SMADi Neural Induction Kit monolayer protocol were differentiated to midbrain-type neural precursors using STEMdiff™ Midbrain Neuron Differentiation Kit for 7 days. (A) Midbrain-type neural precursor cell cultures contain a population of (A) FOXA2-expressing cells (red) and (B) NKX2.1-expressing cells (green). Merge image in (E) showing the bottom half of the same FOV includes DAPI-labeled nuclei (blue) and shows non-overlapping marker expression. The resulting neural precursor cultures also express (C) OTX2 (red) and are negative for (D) central nervous system NPC marker PAX6 (green). Merge image in (F) showing the bottom half of the same FOV includes nuclei that are labeled with DAPI (blue).

Figure 5. Midbrain-Type Neurons Express Dopamine Transporters (DAT) After Differentiation and Maturation in STEMdiff™ Midbrain Neuron Kits

NPCs generated from H9 hPSCs in mTeSR™1 using the STEMdiff™ SMADi Neural Induction Kit monolayer protocol were differentiated and matured to midbrain-type neurons using the STEMdiff™ Midbrain Neuron Differentiation and Maturation Kits. (A) Midbrain-type neurons were formed after NPCs were cultured with the STEMdiff™ Midbrain Neuron Differentiation Kit for 12 days and STEMdiff™ Midbrain Neuron Maturation Kit for 14 days. The resulting cultures contain a population of (B) class III β-tubulin-positive neurons (magenta), which (C) express DAT in blue, and (E) Tyrosine Hydroxylase-positive cells (green). (D) Nuclei are labeled with DAPI (white).