Why 3D cell culture?

Cells grown in vitro are traditionally grown as 2D monolayers, which are easy to set up and maintain. However, these are often mono-cultures (consisting of only one cell type), lack structural architecture due to the absence of extracellular matrix, and are flat with 50% of their surface exposed to tissue culture medium, rather being surrounded by other cells. Hence, 2D cell cultures are a poor approximation of tissues in living organisms.

Developing a simple, highly-consistent and easy-to-use 3D model therefore holds great promise to reduce the number of costly drug failures in clinical trials, provide a more realistic tumour model in oncology, and generally lead to more useful data and more relevant and successful research. There are a range of techniques to grow cells in 3D including spheroids and hydrogels. A scaffold is useful when cell-to-cell and cell-to-matrix contacts are important to cell morphology and behaviour, for cells that do not in nature form spheroids, where more cells are required for sensitive assays than can be grown in a spheroid and where diffusion of gases and proteins is desirable.

HepG2 liver cells in 3D in Mimetix scaffold

HepG2 liver cells were cultivated for 21 days in Mimetix plate, fixed, stained with DAPI and b-actin and imaged with the Nikon Eclipse C1 confocal microscope.

Mimetix® scaffolds for 3D cell culture

  • Mimetix scaffolds are provided in easy-to-use, sterile well plate formats, which are compatible with standard laboratory liquid handling robotics and automated image analysis equipment, requiring minimal protocol adaption when switching from 2D to 3D.
  • The thickness of the scaffolds in Mimetix tissue-culture plates (50 µm) facilitates microscopic imaging in situ while providing the benefits of 3D cell culture to observe 3D cell morphology and behaviour.
  • Mimetix scaffolds have highly consistent architecture, leading to very reproducible results in cell-based assays well-to-well and batch-to-batch, minimising experimental variability.

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Primary Dermal Fibroblasts / Keratinocytes Co-Culture

Primary human dermal fibroblasts and keratinocytes were cultured on our Mimetix scaffold and analysed by fluorescence microsopy after 24 hours and 7 days.

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3D Imaging of Cancer Cells

Confocal fluorescence microscopy is a versatile tool to image cells growing in 3D. This study presents sample images and a Z-stack video of MCF7 breast cancer cells cultured in the Mimetix scaffold.

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Neural Stem Cells

The topology of Electrospinning’s scaffold matrix supports the growth of complex cell types, such as neural stem cells, and their differentiation into mature neurons.

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Hepatotoxicity

HepG2 human liver carcinoma cells are widely-used as a research tool for drug metabolism and toxicity studies. They are easy to culture and readily available in contrast to primary hepatocytes.

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Myelination Assay

Our aligned electrospun Mimetix scaffold allows the differentiation of more than 80% of oligodendrocyte precursors into mature oligodendrocytes in 14 days & the production of sheath lengths comparable to in vivo situation within 7 days by 90% of the oligodendrocytes

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Oncology

There is a significant need for more predictive in vitro efficacy assays in oncology to reduce both the number of costly drug failures in clinical trials and the number of animals used in pre-clinical testing.

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Cardiotoxicity

Cardiomyocytes show a spindle shape with elongated nuclei in the Mimetix scaffold. They respond to antimycin as expected and show almost complete apoptosis in 2h00 (no mitochondrial staining left, annexin positive).