STEMdiff™ 中脑类器官分化试剂盒

用于从人多能干细胞高效生成中脑神经类器官的细胞培养基试剂盒。

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产品号 #100-1096_C

用于从人多能干细胞高效生成中脑神经类器官的细胞培养基试剂盒。

产品优势

采用无基质系统，省去类器官嵌入步骤
在不同细胞系之间获得尺寸一致、形态可重复的类器官
使用 AggreWell™800，每个试剂盒可生成超过 500 个类器官
实现长期培养存活，适用于预测性实验、高通量表型筛选和神经毒性检测
可将模块化区域特异性类器官组合，构建先进的 AssemBloids™

产品组分包括

STEMdiff™ 神经类器官基础培养基 1，20 mL
STEMdiff™ 神经类器官基础培养基 2，500 mL
STEMdiff™ 神经类器官补充剂 A，10 mL
STEMdiff™ 神经类器官补充剂 C，0.25 mL
STEMdiff™ 神经类器官补充剂 K，0.5 mL
STEMdiff™ 神经类器官补充剂 L，0.5 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》。

Anti-Adherence Rinsing Solution
Rinsing solution for cultureware to prevent cell adhesion
Trypan Blue
Reagent for counting viable mammalian cells
Gentle Cell Dissociation Reagent
cGMP, enzyme-free cell dissociation reagent
STEMdiff™ Neural Organoid Maintenance Kit
Cell culture medium kit for long-term maintenance and maturation of patterned neural organoids
Hausser Scientific™ Bright-Line Hemocytomet...
Counting chamber with 2 cover glasses
Ultra-Low Adherent Plate for Suspension Cultu...
Sterile, flat-bottom plate, with ultra-low adherent surface for suspension cultures, with lid; 6-well format
Reversible Strainers
Reversible strainer for filtration of single cells and isolation of cellular aggregates, including embryoid bodies and spheroids
AggreWell™800
Microwell culture plates for easy and reproducible production of embryoid bodies and spheroids
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
D-PBS (Without Ca++ and Mg++)
Dulbecco’s phosphate-buffered saline without calcium and magnesium
Falcon® Serological Pipettes, 10 mL
Sterile polystyrene pipettes for cell culture
Culture Dish, Non-Treated
Sterile, flat-bottom, clear polystyrene non-treated dish with lid; 35, 60, or 100 mm formats
Y-27632 (Dihydrochloride)
RHO/ROCK pathway inhibitor; Inhibits ROCK1 and ROCK2
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

使用 STEMdiff™ 中脑类器官分化试剂盒，无需基质嵌入，即可从人多能干细胞中稳定地生成三维、模式化的脑类器官培养物。与AggreWell™800™搭配使用时，该款无血清细胞培养基可防止类器官融合，并支持每个试剂盒生成超过 500 个类器官，便于进行高统计效能重复实验和更深入的纵向研究。

本试剂盒生成的脑区特异性类器官基于Sergiu Paşca博士的实验方案改编，是具有人类中脑发育特征的三维体外模型，具有代表性的细胞组成和结构组织。这些类器官在不同细胞系之间具有一致的尺寸和可重复的形态，为在功能性三维系统中研究多巴胺能神经元提供了可靠模型。

中脑类器官可与使用STEMdiff™ 背侧前脑类器官分化试剂盒（含额外模式化因子）生成的纹状体类器官结合，形成中脑-纹状体 AssemBloid™ 培养物。类器官可使用STEMdiff™ 神经类器官维持试剂盒进行长期培养（>50 天），以支持预测性分析、高通量表型筛选及神经毒性测试。

亚型
专用培养基

细胞类型
神经细胞，PSC衍生，神经干/祖细胞，多能干细胞

应用
细胞培养，鉴定，分化，功能学筛选，免疫荧光，类器官培养，表型鉴定，球状体培养

品牌
STEMdiff

研究领域
疾病建模，药物发现和毒理检测，神经科学，类器官

制剂类别
无血清

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实验数据

Schematic protocol diagram to generate patterned midbrain organoids from iPSCs

Figure 1. Schematic for the STEMdiff™ Midbrain Organoid Differentiation Kit

hPSC-derived midbrain organoids can be generated in 43 days. Organoids are initially formed in AggreWell™800 plates. After 6 days, the organoids are transferred and cultured in suspension, allowing growth and subsequent patterning to the midbrain. Long-term maintenance and further maturation of midbrain organoids can be achieved using STEMdiff™ Neural Organoid Maintenance Kit, see the Product Information Sheet (PIS) for details. hPSC = human pluripotent stem cell

Midbrain organoids generated with the STEMdiff Midbrain Organoid Kit expressing markers for midbrain floorplate progenitors and mature marginal zone dopaminergic neurons at Day 50

Figure 2. Mature STEMdiff™ Midbrain Organoids Exhibit Brain-Region-Specific Marker Expression and Organization

Representative marker expression in Day 50 organoids produced using STEMdiff™ Midbrain Organoid Kits (left panels) is compared to organoids produced using STEMdiff™ Dorsal Forebrain Organoid Kits. (A) DAPI is used for counterstaining. (B) Midbrain organoids have comparatively less PAX6 expression throughout. Dorsal forebrain organoids exhibit elevated expression of dorsal cortical progenitor marker PAX6, organized radially. (C) Midbrain organoids have elevated expression of floorplate progenitor marker FOXA2 that arranges around pseudoventricular regions (see inset) and (D) marginal zone dopaminergic neuron marker GIRK2 that expands from FOXA2 expressing progenitor zones.

Midbrain organoids generated with the STEMdiff Midbrain Organoid Kit expressing tyrosine hydroxylase at Day 50

Figure 3. STEMdiff™ Midbrain Organoids Express Catecholaminergic Protein Tyrosine Hydroxylase

Midbrain organoids were generated from human ES cell line H7 and iPS cell line STiPS-F016 using STEMdiff™ Midbrain Organoid Differentiation Kit. Organoids were further matured to Day 50 with STEMdiff™ Neural Organoid Maintenance Kit and are compared to organoids generated with STEMdiff™ Dorsal Forebrain Organoid Differentiation Kit. Both guided organoid types generate neurons that express MAP2, but only midbrain organoids express the catecholaminergic neuron specific marker TH. ES cell = embryonic stem cell; iPS cell = induced pluripotent stem cell; TH = tyrosine hydroxylase

The STEMdiff Midbrain Organoid Kit generates midbrain organoids with consistent morphology from multiple cell lines, shown at Day 25 and Day 50

Figure 4. STEMdiff™ Midbrain Organoid Differentiation Kit Supports the Generation of Homogeneous, Reproducible Neural Organoids

(A-F) Representative phase contrast images of midbrain organoids derived from WLS-1C, H7, and STiPS-F016 hPS cell lines using STEMdiff™ Midbrain Organoid Differentiation Kit show uniform size and spherical morphology at Day 25 and Day 50 in vitro. (G) Diameter measurements of midbrain organoids demonstrate reproducibility across different cell lines (mean ± SD; Day 25, n = 21; Day 50, n = 26). hPS cell = human pluripotent stem cell

Midbrain organoids generated with the STEMdiff Midbrain Organoid Kit expressing midbrain floorplate and dopaminergic marginal zone genes

Figure 5. Neural Organoids Generated with STEMdiff™ Midbrain Organoid Kit Express Key Markers of Brain-Region-Specific Patterning

Single organoids differentiated from multiple cell lines were harvested for RNA at Day 25 and Day 50. ΔΔCT (Fold Change) of midbrain organoids is normalized to TBP and respective dorsal forebrain organoid control (A & B) or respective PSC control (C); mean ± SD, n = 3 - 15. (A) Midbrain floorplate precursor markers FOXA2, LMX1A, and EN1 were elevated in midbrain organoids at Day 25. (B) More mature marginal zone dopaminergic markers NURR1, TH, GIRK2, and PITX3 were elevated at Day 50. (C) Dorsal forebrain marker PAX6 and TBR1 expression is downregulated in Day 25 midbrain organoids. PSC = pluripotent stem cell