- 견적 및 주문
- 제품/실험 Q&A
Microglia (미세아교세포)는 뇌 발달, 신경 발생, 시냅스 가소성 및 항상성 유지에 핵심 역할을 하는 뇌의 면역 세포입니다. 특히 뇌세포에서 생성된 찌꺼기 및 죽은신경세포를 제거하는 등 중추신경계 면역계에서 중요한 역할을 담당합니다. 식세포 작용을 통해 beta-amyloid를 제거하여 알츠하이머를 지연시키지만, 이에의해 지나치게 활성화된 microglia는 오히려 알츠하이머나 파킨슨 등 뇌질환을 유발하기도 합니다. iPSC유래 Microglia를 이용하여 in vivo 실험을 대체할 수 있습니다.
|Cat. No.||Product Name||Starting Material|
|ax1666||Human iPSC-derived Microglial Precursors||Cord blood CD34+ cells derived iPSCs|
|ax0666||Human iPSC-derived Microglia (in 96-well plate format)||Cord blood CD34+ cells derived iPSCs|
|Microglia culture media and reagents|
|Cat. No.||Product Name||Quantity|
|ax0660||Microglia Maintenance Medium||100 mL|
Immunocytochemistry of Axol’s iPSC-derived microglia at Day 18 of in culture.
Immunocytochemistry of Human iPSC-derived Microglia, after 18 days in culture, revealed the expression of
IBA-1 and CX3CR1 (a chemokine receptor) as well as the microglial-specific markers TMEM119
(a transmembrane protein), and P2RY12 (purinergic receptor). All 3 markers co-localized strongly with
the myeloid marker IBA-1, confirming identity of brain-resident-like microglia.
A. Microglia and macrophages from the same donor were stained in parallel for the macrophage/microglia
markers IBA-1 (voltage-gated Ca2+ sensor) and CX3CR1 (chemokine receptor) as well as microglial-specific
markers P2RY12 (purinergic receptor) and TMEM119 (transmembrane protein). Expression of P2RY12 and
TMEM119 is either absent or significantly less in macrophages compared to microglia, confirming microglial
identity. x10 Magnification; Scale bar=400um
B. mRNA expression of 6 key microglial signature genes in iPSCs, iPSC-derived monocytes, iPSC-derived
macrophages and iPSC-derived microglia were determined by qRT-PCR. Cells were lysed from starting iPSCs,
Day0 (monocytes), Day14 (macrophages) or Day18 (microglia) prior to RNA isolation followed by RT. CQ1A,
MERTK, P2RY12, GPR34, and TMEM119 are all differentially upregulated in microglia. The disease-associated
marker TREM2 is expressed at roughly constant levels across the myeloid cell lineage. Fold change was
calculated using the ΔΔCT method, with PDGB as an endogenous control and normalisation to iPSC.
N=3 (iPSC), N=2 (macrophage), N=1 (monocyte, microglia), technical replicates were performed in triplicate
(monocyte, macrophage, microglia) or duplicate (iPSC). Error bars are +- S.E.M.
Axol’s iPSC-derived microglia efficiently phagocytose pHrodo green Zymosan particles
A) pHrodo is a fluorogenic dye that dramatically increases in fluorescence as the pH of the surroundings
becomes more acidic, such is the case in the lysosome. pHrodo® Green Zymosan BioParticles®
(Life Technologies) were added at 0.5 mg/mL to day 19-microglia plated at 1x105sup>/cm2, and incubated
for 3hrs at 37oC, 5% CO2. Nuclei were counterstained with a live Hoechst 33342 stain and live cell imaging
was carried out using an on-stage incubator connected to an EVOS Fl Auto imaging system (Life Technologies).
Two activated microglial cells can be seen consuming increasing numbers of BioParticles® over time leading
greater and more intense fluorescence emission. B) and C) Microglia seeded at 1x105/cm2 (B) or 3x105cm2
(C) were pre-treated with 100 ng/mL lipopolysaccharide (LPS) for 1hr (B), 5, 10μM or 30μM Cytochalasin D
(CytoD, phagocytosis inhibitor) or 1μM or 10μM Latrunculin A (Lat-A) for 30mins (B) or 1hr (C) before
adding pHrodo® Green Zymosan BioParticles® at 0.5 mg/mL and incubating for 3hrs at 37oC, 5% CO2.
Lat-A inhibited phagocytosis more strongly than CytoD, in line with Lat-A being a more potent inhibitor
of actin polymerization. Fluorescence was quantified using a SpectraMax iD3 microplate reader (Molecular
Devices), and values were blanked against a no-cell control well. N=4 for both experiments, One-way
ANOVA *P<0.05, **P<0.01, ***P
A and B) Stimulation with TLR agonists LPS (TLR1/2) and R848 (TLR7/8), as well as the cytokine IL1β and
interferon gamma (IFNγ) induces changes in TNFα and IL-6 release.
Microglia plated at 120,000/cm2 were stimulated with increasing concentrations of LPS, R848, IL1β, and IFNγ
for 20hrs. The concentration of TNFα (A) and IL-6 (B) in the harvested supernatant were measured by alphaLISA.
Except for LPS, there is a clear dose-dependent, positive correlation between simulant concentration and amount
of cytokine released. N=1. Data kindly provided by Grünenthal GmbH, Germany.
A. Immunocytochemistry of Human iPSC-derived Microglia co-cultured with Human iPSC-derived cortical
neurons reveals microglia specific maker expression TMEM119 and neuronal marker β-3 Tubulin.
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