Initial culture behaviour of rat blastocysts on selected feeder cell lines

To increase our understanding of rat embryos in culture and to attempt the isolation of blastocyst‐derived cell lines, we examinated the initial growth behaviour of rat blastocysts from four strains of rat on four different feeder cell layers. The feeders used were a continuous cell line of murine e...

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Veröffentlicht in:Molecular reproduction and development 1995-03, Vol.40 (3), p.311-324
Hauptverfasser: Ouhibi, Nadia, Sullivan, Neil F., English, Jennifer, Colledge, William H., Evans, Martin J., Clarke, Neil J.
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container_end_page 324
container_issue 3
container_start_page 311
container_title Molecular reproduction and development
container_volume 40
creator Ouhibi, Nadia
Sullivan, Neil F.
English, Jennifer
Colledge, William H.
Evans, Martin J.
Clarke, Neil J.
description To increase our understanding of rat embryos in culture and to attempt the isolation of blastocyst‐derived cell lines, we examinated the initial growth behaviour of rat blastocysts from four strains of rat on four different feeder cell layers. The feeders used were a continuous cell line of murine embryonic fibroblasts (STO), primary mouse (MEF) or primary rat (REF) embryonic fibroblasts, and a continuous cell line of rat uterine epithelial cells (RUCs). A medium that gave optimum plating efficiencies for murine ES cells was used in the rat embryo culture. Each culture system allowed hatching and attachment of the blastocysts, that is, the behaviour was similar on each feeder and each strain for the first 2 days in culture. Subsequently, there was a rapid differentiation of the Inner Cell Mass (ICM) cells on fibroblastic feeder cell layers (STO > MEF > REF), and this was generally complete after 3–6 days in primary culture. On RUCs, the ICM was found to increase in size without differentiation up to and including day 4 and in some cases longer. Embryo‐derived cells were obtained by disaggregating and passaging ICMs on REF and RUC feeders. Rounded, refractile, and epithelial‐like cells were isolated on REF and colonies of ES‐like cells on the RUCs. The ES‐like cells were positive for expression of alkaline phosphatase and stage‐specific embryonic‐antigen 1. This is an important first step towards the derivation and culture of pluripotent ES cells from the rat. © 1995 Wiley‐Liss, Inc.
doi_str_mv 10.1002/mrd.1080400307
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Segmentation. Gastrulation. Neurulation</topic><topic>Embryo, Mammalian - cytology</topic><topic>Embryology: invertebrates and vertebrates. Teratology</topic><topic>Embryonic and Fetal Development</topic><topic>Epithelial Cells</topic><topic>ES-like cells</topic><topic>Female</topic><topic>Fibroblasts - cytology</topic><topic>Fundamental and applied biological sciences. 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subjects Alkaline Phosphatase - metabolism
Animals
Biological and medical sciences
Blastocyst - cytology
Blastocyst - enzymology
Blastocyst - immunology
Cell Differentiation
Cell Line
Culture Media
Culture Techniques - methods
Early stages. Segmentation. Gastrulation. Neurulation
Embryo, Mammalian - cytology
Embryology: invertebrates and vertebrates. Teratology
Embryonic and Fetal Development
Epithelial Cells
ES-like cells
Female
Fibroblasts - cytology
Fundamental and applied biological sciences. Psychology
ICM
Immunocytochemistry
Lewis X Antigen - metabolism
Mice
Rat embryos
Rats
Rats, Inbred F344
Rats, Inbred WKY
Rats, Sprague-Dawley
Rats, Wistar
Stem Cells - cytology
Uterine cells
Uterus - cytology
title Initial culture behaviour of rat blastocysts on selected feeder cell lines
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