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。
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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。
For differentiation of human ES or iPS cells into hematopoietic progenitor cells
Differentiation kit for the generation of microglia precursors from human ES and iPS cell-derived hematopoietic progenitor cells
Dulbecco's Modified Eagle's Medium/Nutrient Ham's Mixture F-12 (DMEM/F-12) with 15 mM HEPES buffer
Sterile polypropylene conical tubes for use in cell centrifugation and other cell culture applications
Compatible antibodies for purity assessment of isolated cells
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。
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。
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。
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
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.
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.
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.
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.
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种属 | Human |
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配方类别 | Serum-Free |
无血清神经添加物(50X)
提升神经元功能的无血清基础培养基
用于人脑类器官建立与成熟的培养基试剂盒
用于将人 ES 和 iPS 细胞衍生的神经前体细胞分化为神经元前体细胞的分化试剂盒
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