用于将人 ES 细胞或 iPS 细胞分化为红系祖细胞

用于将人 ES 和 iPS 细胞衍生的神经前体细胞分化为神经元前体细胞的分化试剂盒

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产品号 #（选择产品）

产品号 #08600_C

用于将人 ES 和 iPS 细胞衍生的神经前体细胞分化为神经元前体细胞的分化试剂盒

产品优势

支持从 hPSC 衍生的神经元前体细胞高效生成功能性神经元
可产生高纯度的混合兴奋性和抑制性神经元群体（≥ 90％的神经元），并可以长期维持培养
针对使用 STEMdiff™ SMADi 神经诱导试剂盒生成的神经元前体细胞的分化进行了优化
支持神经活性，以获得生理相关结果
实现从多个人胚胎干细胞和诱导多能干细胞系衍生的神经元前体细胞的可重复成熟

产品组分包括

STEMdiff™ 前脑神经元分化基础培养基，80 mL
STEMdiff™ 前脑神经元分化补充剂，20 mL

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总览

STEMdiff™ 前脑神经元分化试剂盒与 STEMdiff™ 前脑神经元成熟试剂盒（目录号 #08605）配套使用，可将源自人多能干细胞的神经前体细胞分化为前脑型（FOXG1 阳性）的混合神经元群体。本试剂盒已针对 STEMdiff™ SMADi 神经诱导试剂盒提供的神经前体细胞进行了优化。使用这些产品生成的神经元可广泛应用于模拟人类神经发育与神经疾病、药物筛选、毒性测试及细胞治疗的验证。

Data Figures

Figure 1. Schematic for the Embryoid Body Protocol

Forebrain-type neural precursors can be generated in 18 - 19 days from hPSC-derived NPCs after selecting neural rosettes from replated embryoid bodies. For the maturation of precursors to forebrain-type neurons, see the PIS. hPSC = human pluripotent stem cell; NPCs = neural progenitor cells; PIS = product information sheet

Figure 2. Schematic for the Monolayer Protocol

Forebrain-type neural precursors can be generated from NPC monolayers derived from embryonic and induced pluripotent stem cells after three single-cell passages. For the maturation of precursors to forebrain-type neurons, see the PIS. NPC = neural progenitor cell; PIS = product information sheet

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

NPCs generated from hPSCs in mTeSR 1™ using the STEMdiff™ SMADi Neural Induction Kit EB protocol were differentiated and matured to forebrain-type neurons using the STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits. (A) Forebrain-type neurons were formed after iPS cell-derived NPCs were cultured with the STEMdiff™ Forebrain Neuron Differentiation Kit for 7 days and STEMdiff™ Forebrain Neuron Maturation Kit for 14 days. The resulting cultures contain a highly pure population of (B) class III β-tubulin-positive neurons (green), with (C) fewer than 10% astrocytes (GFAP-positive cells, red). (D) Nuclei are labeled with DAPI (blue). NPCs = neural progenitor cells; hPSC = human pluripotent stem cell; EB = embryoid body; iPS = induced pluripotent stem

Figure 4. Downstream Differentiation of Neural Progenitor Cells to Neurons Is Possible Using the STEMdiff™ Differentiation and Maturation Kits

(A) NPCs generated from STiPS-R038 hPSCs in mTeSR™1 using the STEMdiff™ SMADi Neural Induction Kit EB protocol were differentiated and matured to cortical neurons using STEMdiff™ Forebrain Neuron Differentiation Kit for 7 days and STEMdiff™ Forebrain Neuron Maturation Kit for 14 days. The resulting cultures contain a highly pure population of (B) class III β-tubulin-positive neurons (green) with less than 10% GFAP-positive astrocytes (not shown). (C) The generated neurons are also positive for FOXG1 expression (red), indicating a forebrain-type identity. (D) Nuclei are labeled with Hoechst (blue). NPCs = neural progenitor cells; hPSC = human pluripotent stem cell

Figure 5. A Mixed Population of Forebrain-Type Cortical Neurons Is Generated Using the STEMdiff™ Differentiation and Maturation Kits

Forebrain-type neurons generated from iPSC-derived NPCs (line AIW002-02) were cultured using the STEMdiff™ Forebrain Neuron Differentiation Kit for 7 days and subsequently matured for the following 6 weeks using STEMdiff™ Forebrain Neuron Maturation Kit. The resulting cultures contain a mixed population of neurons expressing VGLUT1, a glutamatergic marker of excitatory neurons (green), as well as MAP2-positive neurons, indicating mature neurons (magenta). Nuclei are labeled with Hoechst (blue). Data courtesy of Cecilia Rocha, The Neuro's Early Drug Discovery Unit (EDDU), McGill University. iPSC = induced pluripotent stem cell; NPCs = neural progenitor cells

Figure 6. PSC-Derived Astrocytes and Neurons Can Be Co-Cultured to Model Cell-Cell Interactions In Vitro

NPCs generated from the H1 cell line were differentiated to astrocytes using STEMdiff™ Astrocyte Differentiation and Maturation Kits. H9 cell-derived NPCs were differentiated to forebrain-type neurons using STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits. For co-culture, matured astrocytes were seeded onto forebrain neurons that had been in STEMdiff™ Forebrain Neuron Maturation Medium for at least one week. Co-cultures were then switched to STEMdiff™ Forebrain Neuron Maturation Medium the following day and for the remaining co-culture. (A) Neurons cultured alone, following the co-culture feeding schedule, are labeled with DCX (green). (B) DCX-positive neurons (green) and astrocytes (GFAP, red) can be co-cultured for at least 1 - 2 weeks prior to analysis.For a detailed co-culture protocol, please see the Methods Library. NPCs = neural progenitor cells

Figure 7. PSC-Derived Neurons Survive and Mature when Co-Cultured with PSC-Derived Astrocytes

NPCs generated from the STiPS-R038 cell line were differentiated to astrocytes using STEMdiff™ Astrocyte Differentiation and Maturation Kits. STiPS-M001 cell-derived NPCs were differentiated to forebrain-type neurons using STEMdiff™ Forebrain Neuron Differentiation and Maturation Kits. After co-culture for one week, neurons (A) had significantly increased neurite outgrowth as measured on MAP2-positive neurons with the NeuriteTracer plugin for ImageJ (M Pool et al. J Neurosci Methods, 2008) and (B) were more numerous than neurons cultured alone using the same feeding schedule. *, p < 0.05; **, p < 0.01. NPCs = neural progenitor cells

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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