STEMdiff™ Microglia Maturation Kit

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。

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

Functional microglia can be generated in 4-10 days from microglial precursors after 24 days in STEMdiff™ Microglia Differentiation Medium. For the differentiation of microglial precursors from hPSC-derived hematopoietic progenitor cells, 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.