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Two alternative methods for the retrieval of somatic cell populations from the mouse ovary.
Frost, E R; Ford, E A; Taylor, G; Boeing, S; Beckett, E L; Roman, S D; Lovell-Badge, R; McLaughlin, E A; Sutherland, J M.
Afiliação
  • Frost ER; Priority Research Centre for Reproductive Science, Schools of Biomedical Science & Pharmacy and Environmental & Life Sciences, University of Newcastle, Callaghan, NSW, Australia.
  • Ford EA; Hunter Medical Research Institute, New Lambton Heights, NSW, Australia.
  • Taylor G; Stem Cell Biology and Developmental Genetics Lab, The Francis Crick Institute, London, UK.
  • Boeing S; Priority Research Centre for Reproductive Science, Schools of Biomedical Science & Pharmacy and Environmental & Life Sciences, University of Newcastle, Callaghan, NSW, Australia.
  • Beckett EL; Hunter Medical Research Institute, New Lambton Heights, NSW, Australia.
  • Roman SD; Stem Cell Biology and Developmental Genetics Lab, The Francis Crick Institute, London, UK.
  • Lovell-Badge R; Bioinformatics and Biostatistics Facility, The Francis Crick Institute, London, UK.
  • McLaughlin EA; Scientific Computing-Digital Development Team, The Francis Crick Institute, London, UK.
  • Sutherland JM; Hunter Medical Research Institute, New Lambton Heights, NSW, Australia.
Mol Hum Reprod ; 27(6)2021 05 29.
Article em En | MEDLINE | ID: mdl-33973015
Many modern techniques employed to uncover the molecular fundamentals underlying biological processes require dissociated cells as their starting point/substrate. Investigations into ovarian endocrinology or folliculogenesis, therefore, necessitate robust protocols for dissociating the ovary into its constituent cell populations. While in the mouse, methods to obtain individual, mature follicles are well-established, the separation and isolation of single cells of all types from early mouse follicles, including somatic cells, has been more challenging. Herein we present two methods for the isolation of somatic cells in the ovary. These methods are suitable for a range of applications relating to the study of folliculogenesis and mouse ovarian development. First, an enzymatic dissociation utilising collagenase and a temporary, primary cell culture step using neonatal mouse ovaries which yields large quantities of granulosa cells from primordial, activating, and primary follicles. Second, a rapid papain dissociation resulting in a high viability single cell suspension of ovarian somatic cells in less than an hour, which can be applied from embryonic to adult ovarian samples. Collectively these protocols can be applied to a broad array of investigations with unique advantages and benefits pertaining to both.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Coleta de Tecidos e Órgãos Limite: Animals Idioma: En Ano de publicação: 2021 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Coleta de Tecidos e Órgãos Limite: Animals Idioma: En Ano de publicação: 2021 Tipo de documento: Article