STEMdiff™ 星形胶质细胞分化试剂盒

无血清分化试剂盒，用于从人多能干细胞（hPSC）来源的神经前体细胞生成星形胶质细胞前体。

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产品号 #（选择产品）

产品号 #100-0013_C

无血清分化试剂盒，用于从人多能干细胞（hPSC）来源的神经前体细胞生成星形胶质细胞前体。

产品优势

成分明确且无血清
支持高效生成具有功能性的星形胶质细胞
针对使用 STEMdiff™ SMADi 神经诱导试剂盒生成的神经前体细胞进行优化分化
可重复地从多种人胚胎干细胞（ES）和诱导多能干细胞（iPS）系中生成皮层类型的星形胶质细胞前体

产品组分包括

STEMdiff™ 星形胶质细胞分化基础培养基，80 mL
STEMdiff™ 星形胶质细胞分化补充剂，20 mL

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Need a high-quality cell source? Use the hiPSC SCTi003-A (female) or SCTi004-A (male) control lines, manufactured with mTeSR™ Plus.

要查看实验方案所需的所有配套产品，请参阅《实验方案与技术文档》。

STEMdiff™ Neural Rosette Selection Reagent
Enzyme-free reagent for the selective detachment of neural rosettes
STEMdiff™ SMADi Neural Induction Kit
Serum-free medium kit for highly efficient SMAD inhibition-mediated neural induction of human ES and iPS cells
STEMdiff™ Astrocyte Maturation Kit
Maturation kit for the generation of cortical-type astrocytes from human ES and iPS cell-derived astrocyte precursors
Human iPSC-Derived Neural Progenitor Cells
Frozen human neural progenitor cells differentiated from the human induced pluripotent stem cell line, SCTi003-A
DMEM/F-12 with 15 mM HEPES
Dulbecco's Modified Eagle's Medium/Nutrient Ham's Mixture F-12 (DMEM/F-12) with 15 mM HEPES buffer
Labeling Antibodies
Compatible antibodies for purity assessment of isolated cells

总览

实验数据

产品说明书及文档

应用领域

总览

STEMdiff™ 星形胶质细胞分化试剂盒可用于快速且高效地从使用 STEMdiff™ SMADi 神经诱导试剂盒（目录号：08581）获得的人多能干细胞（hPSC）来源的神经前体细胞（NPCs）中生成星形胶质细胞前体。随后，这些星形胶质细胞前体可通过 STEMdiff™ 无血清星形胶质细胞成熟试剂盒（目录号：100-1666）进一步成熟为星形胶质细胞。

采用该无血清系统，最短仅需 7 周即可从 hPSC 生成高度纯化的星形胶质细胞群体（平均 >70% S100B 阳性、>60% GFAP 阳性星形胶质细胞，<15% doublecortin 阳性神经元），且这些细胞可在体外长期培养维持。使用该系列产品衍生的细胞可广泛应用于人类神经发育和疾病模型构建、药物筛选、毒性测试以及细胞治疗验证等研究领域。

亚型
专用培养基

细胞类型
神经细胞，PSC衍生，神经干/祖细胞

种属
人

应用
细胞培养，分化

品牌
STEMdiff

研究领域
疾病建模，药物发现和毒理检测，神经科学

制剂类别
无血清

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实验数据

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

Figure 1. Schematic for the Embryoid Body Protocol

Cortical-type astrocyte precursors can be generated in 20 days from hPSC-derived neural progenitor cells (NPCs) after selecting neural rosettes from replated embryoid bodies. For the maturation of precursors to cortical-type astrocytes, see the PIS.

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

Figure 2. Schematic for the Monolayer Protocol

Cortical-type astrocyte precursors can be generated in 21 days 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 cortical-type astrocytes, see the PIS.

Culturing PSCs in STEMdiff™ SMADi Neural Induction Kit and STEMdiff™ Astrocyte Differentiation and Maturation Kits Yields Cortical-Type Astrocytes

Figure 3. Cortical-Type Astrocytes Are Generated After Culture in STEMdiff™ Astrocyte Differentiation and Maturation Kits

NPCs generated from hPSCs in TeSR™-E8™ using the STEMdiff™ SMADi Neural Induction Kit embryoid body (EB) protocol were differentiated and matured to cortical-type astrocytes using the STEMdiff™ Astrocyte Differentiation and Maturation Kits. Cortical-type astrocytes were formed after iPS cell-derived NPCs were cultured with the STEMdiff™ Astrocyte Differentiation Kit for 3 weeks and STEMdiff™ Astrocyte Maturation Kit for 3 weeks. (A) Nuclei are labeled with DAPI (gray). The resulting cultures contain a highly pure population of astrocytes, which are (B) more than 60% GFAP-positive (green) and (C) more than 70% S100B-positive (magenta), with (D) fewer than 15% neurons (DCX-positive cells, cyan). Scale bar = 100 μm.

Figure 4. STEMdiff™ Astrocyte Kits Generate Cells Expressing Expected Levels of Genes Characteristic for Astrocytes

Embryonic stem and induced pluripotent stem cells from a variety of lines (n = 6, maintained in mTeSR™1 or TeSR™-E8™) were differentiated to NPCs using the STEMdiff™ SMADi Neural Induction Kit embryoid body protocol. Cells were then grown in STEMdiff™ Astrocyte Differentiation Kit for 3 weeks followed by STEMdiff™ Astrocyte Maturation Kit for 3 weeks prior to analysis. Expression levels were measured by quantitative PCR (qPCR) and normalized to hPSC controls relative to housekeeping genes 18S and TBP.

Figure 5. 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.

Figure 6. 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