STEMdiff™ 小胶质细胞成熟试剂盒

用于将人胚胎干细胞（ES）和诱导多能干细胞（iPS）来源的小胶质细胞前体分化成熟为小胶质细胞的成熟试剂盒。

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产品号 #（选择产品）

产品号 #100-0020_C

用于将人胚胎干细胞（ES）和诱导多能干细胞（iPS）来源的小胶质细胞前体分化成熟为小胶质细胞的成熟试剂盒。

产品优势

成分明确、无血清的体系，以获得一致且可重复的结果。
将超过 90% 的造血祖细胞（HPCs）分化为高纯度的小胶质细胞。
生成具有吞噬和激活功能的小胶质细胞。
每个试剂盒平均可实现功能性小胶质细胞的四倍扩增。
与 STEMdiff™ 神经类器官共培养体系兼容。

产品组分包括

STEMdiff™ 小胶质细胞成熟试剂盒（目录号 #100-0020）

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
STEMdiff™ 小胶质细胞补充剂 3，400 μL

立即询价

总览

STEMdiff™ 小胶质细胞培养系统包括 STEMdiff™ 小胶质细胞分化试剂盒和 STEMdiff™ 小胶质细胞成熟试剂盒。这两种试剂盒结合使用，用于分化和成熟采用 STEMdiff™ 造血试剂盒（货号 #05310）获得的人类多能干细胞 (hPSC) 衍生小胶质细胞。
该系统基于 Mathew Blurton-Jones 实验室的方案（Abud 等，2017），生成的细胞为高纯度的小胶质细胞群（>80% CD45/CD11b 阳性，>50% TREM2 阳性小胶质细胞；形态上明显不同的单核细胞或巨噬细胞占比 <20%）。

使用该系列产品获得的细胞可作为研究神经炎症、人类神经系统发育与疾病、共培养实验以及毒性测试的多功能工具。

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.

Microglia Generated Using the STEMdiff™ Microglia Culture System Exhibit Robust Expansion, Mature Phenotypic Markers, and Homeostatic Morphology

Figure 2. Microglia Generated Using the STEMdiff™ Microglia Culture System Exhibit Robust Expansion, Mature Phenotypic Markers, and Homeostatic Morphology

(A) Microglia generated using the STEMdiff™ Microglia Culture System undergo a four-fold expansion, on average, across four cell lines. The fold expansion was calculated by taking the total cell count at Day 24 and dividing it by the number of seeded cells at Day 0. The bars show the mean ± standard deviation. Technical replicates were averaged, n = 1 - 4 technical replicates, 1 - 9 experimental setups.

(B) Microglia generated with STEMdiff™ Microglia Culture System have CD45+ CD11b+ co-expression and P2RY12+ expression as measured by flow cytometry on Day 24. The bars show the mean ± standard deviation.Technical replicates (n = 1 - 4) were averaged, and each dot in the graph represents an experimental replicate.

(C) Normal microglial morphology, characterized by small cell bodies and ramified processes, is observed in cells generated using the STEMdiff™ Microglia Culture System. Images at Days 12 and 24 were captured prior to replate and harvest. Scale bar = 100 µm.

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

Figure 3. 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 4. 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.

Figure 5. STEMdiff™ Microglia Culture System Generates Functional Microglia Capable of Phagocytosis at Day 34

Microglia taking up pH-sensitive bioindicator particles at a concentration of 5 μg/mL were measured over a 72-hour time period with live cell imaging. As the particles are phagocytosed, the particles turn red and are concentrated within the cells. Over time, the microglia display an activated ameboid morphology. Scale bar = 400 μm.

Figure 6. 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.

产品说明书及文档

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