A three-dimensional collagen construct to model lipopolysaccharide-induced activation of BV2 microglia

Journal of Neuroinflammation

Sharmili Vidyadaran³  James B Phillips¹  Han Chung Lee²  Andrew Yew³  Chih Kong Tong³  Randy Tatt Yhew Haw³ 

[1] Department of Biomaterials & Tissue Engineering, University College London, UCL Eastman Dental Institute, 256 Gray’s Inn Road, London WC1X 8LD, UK;Genetic & Regenerative Medicine Research Centre (GRMRC) & Department of Obstetrics & Gynaecology, Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia;Neuroinflammation Group, Immunology Laboratory, Department of Pathology, Faculty of Medicine & Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

关键词: Three-dimensional cultures;    Collagen matrix;    Lipopolysaccharide;    Microglia;   

Others  :  1151203
DOI  :  10.1186/1742-2094-11-134

 received in 2013-12-16, accepted in 2014-07-16,  发布年份 2014

【 摘 要 】

Background

We report a novel method of culturing microglia in three dimension (3D) using collagen as a substrate. By culturing microglia within a matrix, we aim to emulate the physical state of microglia embedded within parenchyma.

Methods

BV2 microglia cell suspensions were prepared with type I collagen and cast into culture plates. To characterise the BV2 microglia cultured in 3D, the cultures were evaluated for their viability, cell morphology and response to lipopolysaccharide (LPS) activation. Conventional monolayer cultures (grown on uncoated and collagen-coated polystyrene) were set up concurrently for comparison.

Results

BV2 microglia in 3D collagen matrices were viable at 48 hrs of culture and exhibit a ramified morphology with multiplanar cytoplasmic projections. Following stimulation with 1 μg/ml LPS, microglia cultured in 3D collagen gels increase their expression of nitric oxide (NO) and CD40, indicating their capacity to become activated within the matrix. Up to 97.8% of BV2 microglia grown in 3D cultures gained CD40 positivity in response to LPS, compared to approximately 60% of cells grown in a monolayer (P < .05). BV2 microglia in 3D collagen gels also showed increased mRNA and protein expression of inflammatory cytokines IL-6, TNF-α and the chemoattractant MCP-1 following LPS stimulation.

Conclusions

In summary, BV2 microglia cultured in 3D collagen hydrogels exhibit multiplanar cytoplasmic projections and undergo a characteristic and robust activation response to LPS. This culture system is accessible to a wide range of analyses and provides a useful new in vitro tool for research into microglial activation.

【 授权许可】

   
2014 Haw et al.; licensee BioMed Central Ltd.

【 图 表 】

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