Isolation and culturing of zebrafish pluripotent cells

2.50
Hdl Handle:
http://hdl.handle.net/10547/347064
Title:
Isolation and culturing of zebrafish pluripotent cells
Authors:
Sana, Salma
Abstract:
Zebrafish (Dania rerio) is an important model organIsm for the studies of vertebrate development and gene expression in the field of molecular biology and biomedicine. Its embryonic stem cells provide a unique tool linking in vitra and in vivo genetic manipulations of animal genomes. The aim of the project was to determine the most suitable embryo stage for the isolation and culturing of pluripotent cells of zebrafish embryos. Studies were carried out to investigate the expression of two pluripotency markers i.e. Oct4 and Sa1l4 at certain embryonic stages employing Immunohistochemistry. The protein expression studies indicated maximum expression of Oct4 and Sa1l4 at high stage. Primary cultures were initiated from zebrafish high stage embryos in basal nutrient medium; supplemented with insulin, selenite, epidermal growth factor and Foetal Bovine serum. Experiments were conducted for the determination of optimum concentrations of FBS. The growing cultures identified the signs of differentiation i.e. melanocytes and neurite formation. Basic fibroblast growth factor (bFGF) was found to inhibit the differentiation in cultures. The developed pluripotency markers were tested on cultured embryonic cells. Results indicated that these markers work well for the identification ofphenotype of embryonic cell colonies in zebrafish. The establishment of pluripotent cell line will enable knock out and knock in technologies to be used in this species and have important applications in functional genomics research.
Citation:
Sana, S. (2012) 'Isolation and culturing of zebrafish pluripotent cells'. Masters by Research thesis. University of Bedfordshire.
Publisher:
University of Bedfordshire
Issue Date:
Oct-2012
URI:
http://hdl.handle.net/10547/347064
Type:
Thesis or dissertation
Language:
en
Description:
A thesis submitted to the University of Bedfordshire in partial fulfilment of the requirements for the degree of Masters by Research
Appears in Collections:
Masters e-theses

Full metadata record

DC FieldValue Language
dc.contributor.authorSana, Salmaen
dc.date.accessioned2015-03-24T12:36:08Zen
dc.date.available2015-03-24T12:36:08Zen
dc.date.issued2012-10en
dc.identifier.citationSana, S. (2012) 'Isolation and culturing of zebrafish pluripotent cells'. Masters by Research thesis. University of Bedfordshire.en
dc.identifier.urihttp://hdl.handle.net/10547/347064en
dc.descriptionA thesis submitted to the University of Bedfordshire in partial fulfilment of the requirements for the degree of Masters by Researchen
dc.description.abstractZebrafish (Dania rerio) is an important model organIsm for the studies of vertebrate development and gene expression in the field of molecular biology and biomedicine. Its embryonic stem cells provide a unique tool linking in vitra and in vivo genetic manipulations of animal genomes. The aim of the project was to determine the most suitable embryo stage for the isolation and culturing of pluripotent cells of zebrafish embryos. Studies were carried out to investigate the expression of two pluripotency markers i.e. Oct4 and Sa1l4 at certain embryonic stages employing Immunohistochemistry. The protein expression studies indicated maximum expression of Oct4 and Sa1l4 at high stage. Primary cultures were initiated from zebrafish high stage embryos in basal nutrient medium; supplemented with insulin, selenite, epidermal growth factor and Foetal Bovine serum. Experiments were conducted for the determination of optimum concentrations of FBS. The growing cultures identified the signs of differentiation i.e. melanocytes and neurite formation. Basic fibroblast growth factor (bFGF) was found to inhibit the differentiation in cultures. The developed pluripotency markers were tested on cultured embryonic cells. Results indicated that these markers work well for the identification ofphenotype of embryonic cell colonies in zebrafish. The establishment of pluripotent cell line will enable knock out and knock in technologies to be used in this species and have important applications in functional genomics research.en
dc.language.isoenen
dc.publisherUniversity of Bedfordshireen
dc.subjectC410 Applied Geneticsen
dc.subjectpluripotent cellsen
dc.subjectzebrafishen
dc.subjectDanio rerioen
dc.subjectembryosen
dc.titleIsolation and culturing of zebrafish pluripotent cellsen
dc.typeThesis or dissertationen
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