STEMdiff™ 前脑神经元成熟试剂盒

用于从人ES和iPS细胞衍生的神经元前体细胞生成功能性神经元的成熟试剂盒

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产品号 #（选择产品）

产品号 #08605_C

用于从人ES和iPS细胞衍生的神经元前体细胞生成功能性神经元的成熟试剂盒

产品优势

成分明确且无血清
支持从 hPSC 衍生的神经元前体细胞高效生成功能性神经元
生成高纯度的混合兴奋性和抑制性神经元群体（≥ 90％的神经元），可能够在培养中长期维持
针对使用 STEMdiff™ 前脑神经元分化试剂盒生成的神经元前体细胞的成熟进行了优化
支持神经活动，以获得生理相关结果
能够实现多个人胚胎干细胞和诱导多能干细胞系衍生的神经前体细胞的可重复成熟

产品组分包括

BrainPhys™ 神经元培养基，100 mL
STEMdiff™ 前脑神经元成熟补充剂，25 mL

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总览

STEMdiff™ 前脑神经元成熟试剂盒与 STEMdiff™ 前脑神经元分化试剂盒（目录号 #08600）配合使用，可从源自人类多能干细胞的神经祖细胞中生成混合的前脑型（FOXG1 阳性）神经元群。所获得的神经元纯度高（III 类 β-微管蛋白阳性神经元≥ 90%；GFAP 阳性星形胶质细胞< 10%），功能齐全，可在培养中长期维持。使用这些产品获得的神经元是构建人类神经发育和疾病模型、药物筛选、毒性测试和细胞疗法验证的多功能工具。

实验数据

Figure 1. Schematic for the Embryoid Body Protocol

Forebrain-type neurons can be generated from NPCs after 6 days in STEMdiff™ Forebrain Neuron Differentiation Medium. For differentiation of precursors from embryonic and induced pluripotent stem cells, see the PIS. NPCs = neural progenitor cells; PIS = product information sheet

Figure 2. Schematic for the Monolayer Protocol

Forebrain-type neurons can be generated from NPCs after 6 days in STEMdiff™ Forebrain Neuron Differentiation Medium. For differentiation of precursors from embryonic and induced pluripotent stem cells, see the PIS. NPC = neural progenitor cells; 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

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