STEMdiff™ 小胶质细胞分化试剂盒

用于从人ES和iPS细胞衍生的造血祖细胞生成小胶质细胞前体的分化试剂盒

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产品号 #（选择产品）

产品号 #100-0019_C

用于从人ES和iPS细胞衍生的造血祖细胞生成小胶质细胞前体的分化试剂盒

产品优势

超过90%的来源HPC成功分化为小胶质细胞。
与使用 STEMdiff™ 造血试剂盒生成的 HPC 具有上游兼容性，并与 STEMdiff™ 神经类器官共培养。
可实现小胶质细胞的分化，且几乎没有巨噬细胞或单核细胞污染。
使用本试剂盒生成的小胶质细胞具有吞噬和激活功能。
所需材料以简单、易于使用的格式提供。

产品组分包括

STEMdiff™ 小胶质细胞分化试剂盒（目录号100-0019）

STEMdiff™ 小胶质细胞基础培养基，90 mL
STEMdiff™ 小胶质细胞补充剂 1，10 mL
STEMdiff™ 小胶质细胞补充剂 2，400 μL

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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™ Hematopoietic Kit
For differentiation of human ES or iPS cells into hematopoietic progenitor cells
STEMdiff™ Microglia Maturation Kit
Maturation kit for the generation of microglia from human ES and iPS cell-derived microglia precursors
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
Falcon® Conical Tubes, 15 mL
Sterile polypropylene conical tubes for use in cell centrifugation and other cell culture applications
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™ 小胶质细胞培养系统包含 STEMdiff™ 小胶质细胞分化试剂盒和 STEMdiff™ 小胶质细胞成熟试剂盒。该系统与 STEMdiff™ 造血试剂盒（目录号：05310）配合使用，可用于将人多能干细胞（hPSCs）分化并成熟为小胶质细胞。

根据 Mathew Blurton-Jones 实验室 (Abud et al., 2017) 的方案，所得细胞为高纯度的小胶质细胞群（CD45/CD11b 阳性细胞比例 > 80%，TREM2 阳性细胞比例 > 50%；形态学可识别的的单核细胞或巨噬细胞比例 < 20%）。

本系统基于 Mathew Blurton-Jones 实验室的实验方案（Abud 等，2017），所生成的细胞为高纯度的小胶质细胞群（CD45/CD11b 阳性细胞比例 > 80%，TREM2 阳性细胞比例 > 50%；形态学明显不同的单核细胞或巨噬细胞比例 < 20%）。。

使用这些产品获得的细胞可作为神经炎症建模、人类神经发育和疾病研究、共培养应用以及毒性测试的多功能工具。

亚型
专用培养基

细胞类型
造血细胞，PSC衍生，小胶质细胞，神经细胞，PSC衍生

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，免疫，神经科学

制剂类别
无血清

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实验数据

Experimental protocol schematic for generating microglial precursors from hPSC-derived hematopoietic progenitor cells.

Figure 1. Schematic for the STEMdiff™ Microglia Culture System Protocol

Microglial precursors can be generated in 24 days from hPSC-derived hematopoietic progenitor cells. For the generation of hematopoietic progenitor cells, see documentation for STEMdiff™ Hematopoietic Kit (Catalog #05310). For the maturation of microglial precursors to functional microglia, see the PIS.

Bulk RNA-seq heatmap showing that STEMdiff™ Microglia express disease-relevant genes similar to those from popular protocols.

Figure 2. Microglia Generated with STEMdiff™ Microglia Culture System Express Disease-Relevant Genes Similar to Those from Published Differentiation and Maturation Protocols

Bulk RNA-seq datasets were extracted from 8 different publications that generated hPSC- (iMGL) and primary- (MGL) derived microglia and their transcriptional profiles compared to data from microglia generated with STEMdiff™ Microglia Culture System. The heat map displays absolute expression levels for select genes associated with Alzheimer’s disease, Parkinson’s disease, and viral encephalitis. Significant differences in gene expression between microglia generated with STEMdiff™ Microglia Culture System and any of the other 3 groups were identified by differential gene expression analysis. *= p<0.05 (DEseq2, adjusted). hPSC = human pluripotent stem cell.

Graphs demonstrating that STEMdiff™ Microglia release cytokines in response to LPS stimulation, compared to control.

Figure 3. Microglia Generated with STEMdiff™ Microglia Culture System Release Cytokines in Response to Inflammatory Signals

Microglia were generated using the STEMdiff™ Microglia Culture System and stimulated with 100 ng/mL LPS for 24 hours. The release of pro-inflammatory (TNFα, IL-6, IFN-γ, IL-1β, GM-CSF, IL-12p70, IL-2, IL-8) and anti-inflammatory (IL-10) cytokines were measured by MSD. The microglia release cytokines in response to LPS treatment, as expected. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001. LPS = lipopolysaccharide; MSD = Meso Scale Discovery.

Time lapse of functional microglia phagocytosing bioindicator particles in culture. Over time, cells turn red and aggregate.

Figure 4. STEMdiff™ Microglia Culture System Generates Functional Microglia Capable of Phagocytosis at Day 34

Microglia taking up pH-sensitive bioindicator particles at a concentration of 250 μg/mL (small dots) were measured over a 12-hour time period with live cell imaging. As the particles are phagocytosed, the cells turn red. Over time, the number of small dots decreased, and the red cells increased in number and aggregated. Scale bar = 100μm.

Figure 5. PSC-Derived Microglia Incorporate into Brain Organoids After 10 Days and Display an Activated Morphology upon Injury.

(A) Representative microglia and brain organoid co-cultures after 10 days, stained with IBA1 for microglia (green) and MAP2 for neurons (magenta). The microglia integrate among the neurons and display an unactivated morphology with extended processes (arrow). (B) The microglia display an activated amoeboid morphology upon injury as shown by IBA1 staining.