Challenges in Microglia Research? Unlock Key Technical Solutions with This Guide

I. In Vitro Research: 4 Types of Cell Model Solutions

1. Primary Cells

Application Scenarios: Mechanistic research requiring maximum retention of in vivo characteristics;

Breakthrough: Human/mouse brain tissue sorting technology (preserves original biological properties);

Notes: Strict aseptic operation is required to avoid phenotypic drift caused by long culture cycles.

2. Immortalized Cell Lines

Recommended Cells: Human-derived HMC3, HMO6, and mouse-derived BV-2, N9, etc.;

Core Advantages: Low cost, short cycle, suitable for high-throughput drug screening;

Limitation Tips: Be vigilant against genetic variation during long-term passage.

3. Induced Microglia-Like Cells

Technical Breakthrough: iPSC differentiation technology + Amyloid β-Protein stimulation (abs45128173, ANT BIO PTE. LTD.) = successful simulation of disease-associated phenotypes (DAM);

Application Value: A gold standard tool for studying the mechanisms of genetic diseases.

4. Brain Organoids

Cutting-Edge Protocol: Overexpression of PU.1 induces the generation of functional microglia in cortical organoids. Further xenotransplantation of the organoid model into mice allows vascularized reconstructed organoids to provide in vivo-like nutrition and signal support for the development of microglia.

Model Advantages: Enables the study of cell-cell interactions in a 3D structure, provides a physiological-like research platform, and is suitable for disease model construction.

II. In Vivo Research: 3 Major Animal Model Solutions

1. Drug Depletion Model[1]

The CSF1R small-molecule inhibitor PLX5622 (abs823427, ANT BIO PTE. LTD.) achieves efficient depletion by targeting CSF1R with its advantage of high blood-brain barrier penetration. Among them, administration of PLX5622 for 3 days can achieve an 80% cell depletion rate, and the depletion effect can last for up to 6 months. It has both high specificity and low inflammation risk, and has become the gold standard for microglia depletion in mice.

2. Aging Research Model[2]

3-DR Model: Three rounds of depletion with PLX5622 (abs823427, ANT BIO PTE. LTD.) → targeted simulation of aging status

Aging Mechanism: Three rounds of forced proliferation (>20 divisions) → telomere loss/DNA damage accumulation → permanent senescence

1st Round of Administration: PLX5622 treatment for 7 days, microglia depletion >90%, proliferation again 7 days after drug withdrawal; 2nd Round of Administration: Re-depletion + re-proliferation; 3rd Round of Administration: Induction of cellular replicative senescence

Compared with natural aging, the cycle is shortened by 80%, avoiding the interference of synchronous aging of multiple cells.

3. Transgenic Models

Microglia gene editing technologies mainly include the Cre-loxP recombinase system, CRISPR/Cas gene editing technology, and viral transduction technology.

III. In Vivo Imaging Technology of Microglia

Near-infrared window II (NIR-II, 1000-1700 nm) cyanine dyes provide a breakthrough tool for in vivo imaging of microglia due to their advantages of high tissue penetration, low background interference, and high spatiotemporal resolution.

Targeted Probe NIR-II Imaging

Probe Design: Coupling cyanine dyes such as ICG (abs816408, ANT BIO PTE. LTD.), IR-820 (abs825392, ANT BIO PTE. LTD.) with ligands (such as antibodies, peptides) targeting microglia surface markers such as TREM2 (abs05663, ANT BIO PTE. LTD.), CSF1R (abs06312, ANT BIO PTE. LTD.) to construct specific probes (such as HSA-ICG-iRGD as in similar literatures, Figure 1).

Figure 1 Schematic diagram of HSA-ICG-iRGD for active-targeted NIR-II imaging and photothermal therapy in orthotopic glioma mice[3]

0, 2 min, 3h, 12h, 24h, Ex.vivo; J: 0, 8, 14, 11, 17 days; G1: tumor, PBS; G2: PBS + Laser; G3: HSA-ICG; G4: HSA-ICG + Laser; G5: HSA-ICG-iRGD; HSA-ICG-iRGD + Laser

Imaging Advantages:

•         Penetration depth up to centimeter level (3-5 times improvement in brain tissue penetration depth), overcoming skull occlusion;

•         Signal-to-background ratio (SBR) > 6 (literature cases reach 6.85), clearly distinguishing activated/resting microglia;

•         Dynamic monitoring of microglia migration (e.g., aggregation process towards Aβ plaques).

Application Scenarios: Real-time tracking of microglia phagocytosis of Aβ in Alzheimer's disease models.

From precise depletion and dynamic imaging to disease modeling, the bottlenecks in microglia research are being systematically broken! Whether it is the gold standard of three-day efficient depletion with PLX5622, the in vivo monitoring capability of NIR-II probes penetrating the skull, or the pathological model construction with iPSC differentiation + organoid construction, these technological innovations are pushing the research on the mechanisms of neurological diseases into a new dimension.

References:

[1] Boland R, Kokiko-Cochran ON. Deplete and repeat: Microglial CSF1R inhibition and traumatic brain injury. Front Cell Neurosci. 2024,18:1352790.

[2] Li X, Li Y, Jin Y, et al. Transcriptional and epigenetic decoding of the microglial aging process.Nat Aging. 2023,3(10):1288~1311.

[3] Wu Y Y, Hu D H, Gao D Y, et al. Miniature NIR-Ⅱ nanoprobes for active-targeted phototheranostics of brain tumors[J]. Advanced Healthcare Materials, 2022, 11(23): e2202379.

Product Recommendations for This Issue:

Product Recommendations for Neurological Disease Model Induction:

Brand Mission

ANT BIO PTE. LTD. is dedicated to advancing life science research by providing high-quality, reliable reagents and comprehensive solutions. We recognize the critical role of microglia research in unraveling the mechanisms of neurological diseases and the urgent need for innovative technical tools to accelerate research progress. Through our specialized sub-brands (Absin, Starter, UA), we have developed a full-spectrum product portfolio tailored to microglia research needs, covering cell model induction reagents, animal model construction tools, in vivo imaging probes, and specific proteins.

Our team adheres to stringent quality control standards throughout the product development and production process, ensuring the consistency, purity, and reliability of each product. We are committed to providing professional technical support and customer-centric services, helping researchers overcome experimental challenges such as phenotypic drift and low imaging resolution, and accelerate the pace of microglia research breakthroughs. ANT BIO PTE. LTD. strives to be a trusted partner for scientists worldwide, contributing to the advancement of neuroscience research and the development of therapeutic strategies for neurological diseases.

Disclaimer

This article is AI-compiled and interpreted based on the original work related to microglia research. All intellectual property (e.g., images, data) of the original publication shall belong to the journal and the research team. For any infringement, please contact us promptly and we will take immediate action.

Brand Promotion Copy

ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs

At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.