Seeding hESCs to achieve optimal colony clonality

Wadkin, L. E., Orozco-Fuentes, Sirio, Neganova, I., Bojic, S., Laude, A., Lako, M., Parker, N. G. and Shukurov, A. (2019) Seeding hESCs to achieve optimal colony clonality. Scientific Reports, 9 (1). p. 15299. ISSN 2045-2322

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Official URL: https://doi.org/10.1038/s41598-019-51897-0

Abstract

Human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) have promising clinical applications which often rely on clonally-homogeneous cell populations. To achieve this, it is important to ensure that each colony originates from a single founding cell and to avoid subsequent merging of colonies during their growth. Clonal homogeneity can be obtained with low seeding densities; however, this leads to low yield and viability. It is therefore important to quantitatively assess how seeding density affects clonality loss so that experimental protocols can be optimised to meet the required standards. Here we develop a quantitative framework for modelling the growth of hESC colonies from a given seeding density based on stochastic exponential growth. This allows us to identify the timescales for colony merges and over which colony size no longer predicts the number of founding cells. We demonstrate the success of our model by applying it to our own experiments of hESC colony growth; while this is based on a particular experimental set-up, the model can be applied more generally to other cell lines and experimental conditions to predict these important timescales.

Item Type: Article
Uncontrolled Keywords: Applied mathematics, Embryonic stem cells
Subjects: F300 Physics
G100 Mathematics
Department: Faculties > Engineering and Environment > Mathematics, Physics and Electrical Engineering
Depositing User: Rachel Branson
Date Deposited: 06 Apr 2020 12:37
Last Modified: 06 Apr 2020 12:45
URI: http://nrl.northumbria.ac.uk/id/eprint/42702

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