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Methodological Insights on Morphometric Comparison of Collagen-Type-1 and Polystyrene Grown Malignant Glioma Cells Upon Chemical Stress Induction

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Proceedings of the National Academy of Sciences, India Section B: Biological Sciences Aims and scope Submit manuscript

Abstract

Lack of arrangement complexity in cells grown on 2D cultures for in-vitro experiments has been shown to be a major contributor to lack of reproducibility of in-vivo experiments. Culturing cells along with extracellular matrix (ECM) materials like Collagen has shown promise towards bridging this gap. Cells grown with ECM materials, display entirely different morphological behavior from their polystyrene plate based counterparts. Being commercially expensive, protocols for efficient, in-house collagen-1 extraction for cell culture purpose have been detailed in literatures. Present work focuses on the methods of quantifying and detailing the nuclear morphology (area, perimeter, circularity, roundness, solidity, AR (aspect-ratio) and fractal dimension) of cells grown on Collagen type-1, extracted in-house from rat tails. Treatment of cells grown on collagen and normal polystyrene base plates with established chemical stressors (Acetaminophen, Cobalt chloride and Hydrogen peroxide) was done to demonstrate morphometric differences in the respective conditions. Significant changes were observed in the nuclear morphology of gel grown cells. Application of stress factors in many cases led cells to exhibit morphological adjustments favoring their invasion into the Collagen gel underneath. These changes were in contrast to the polystyrene cultures, which mostly exhibited behaviors showing nuclear flattening and swelling. 3D cell cultures have the potential to produce more accurate experimental results than their 2D counterparts, increasing their reproducibility in further in-vivo tests.

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Acknowledgements

The authors would like to acknowledge Ms. Pearlin (Ph.D research fellow- CBCMT) for helping us source the Rat tails used to extract Collagen in the present work and for her constant guidance. The authors also thank the CBCMT members for their valuable critical comments.

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Authors and Affiliations

  1. Center for Biomaterials, Cellular and Molecular Theranostics (CBCMT), VIT, Vellore, India

    Jishnu Nair & Raunak Kumar Das

  2. School of Bio-Sciences and Technology (SBST), VIT, Vellore, India

    Jishnu Nair

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  1. Jishnu Nair
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Correspondence to Raunak Kumar Das.

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Significance statement:

A methodological insight into quantification and comparison of cellular morphological aspects upon culture in a Collagen based gel and polystyrene base plates is provided. Changes in these aspects with stress induction is also detailed.

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Nair, J., Das, R.K. Methodological Insights on Morphometric Comparison of Collagen-Type-1 and Polystyrene Grown Malignant Glioma Cells Upon Chemical Stress Induction. Proc. Natl. Acad. Sci., India, Sect. B Biol. Sci. 94, 301–314 (2024). https://doi.org/10.1007/s40011-023-01528-6

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