Seeding hESCs to achieve optimal colony clonality.
Sci Rep
; 9(1): 15299, 2019 10 25.
Article
en En
| MEDLINE
| ID: mdl-31653933
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.
Texto completo:
1
Base de datos:
MEDLINE
Asunto principal:
Diferenciación Celular
/
Técnicas de Cultivo de Célula
/
Proliferación Celular
/
Células Madre Pluripotentes Inducidas
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Células Madre Embrionarias Humanas
Tipo de estudio:
Guideline
/
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Sci Rep
Año:
2019
Tipo del documento:
Article
País de afiliación:
Reino Unido