Embryonic Organ Culture

Takayoshi Sakai1, Tomohiro Onodera1

1 Laboratory of Cell and Developmental Biology, National Institute of Dental and Craniofacial Research, National Institutes of Health, Bethesda, Maryland
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 19.8
DOI:  10.1002/0471143030.cb1908s41
Online Posting Date:  December, 2008
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Abstract

This unit provides detailed protocols for dissecting embryonic organs and performing organ culture to study questions in developmental biology. Procedures are described here for dissecting organs such as kidney, lung, and salivary gland. The unit also contains commentary including troubleshooting for embryonic organ culture. Curr. Protoc. Cell Biol. 41:19.8.1‐19.8.8. © 2008 by John Wiley & Sons, Inc.

Keywords: embryonic organ culture; salivary gland; lung; kidney; branching morphogenesis

     
 
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Table of Contents

  • Introduction
  • Basic Protocol 1: Mouse Dissection
  • Basic Protocol 2: Organ Culture
  • Reagents And solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Mouse Dissection

  Materials
  • 70% ethanol
  • DMEM/F12 medium (Invitrogen), sterile
  • Pregnant ICR strain mice: embryonic day 12 (E12) to day 14 (E14)
  • Dissecting instruments including:
    • Curved forceps, length 120 mm
    • Two straight dissecting forceps, length 110 mm
    • Straight scissors, length 140 mm
    • Scalpel blade, size no. 11
    • Glass dissection plate
  • 100‐mm and 30‐mm culture dishes, sterile
  • Glass dissection plate, 10 to 15 cm, square or circular
  • Dissection microscope
  • Additional reagents and equipment for euthanasia of the mouse (Donovan and Brown, )

Basic Protocol 2: Organ Culture

  Materials
  • Culture medium (see recipe)
  • Mouse embryonic organs ( protocol 1)
  • 50‐mm glass‐bottom dish (MatTek Corporation)
  • Filter membrane (diameter 13 mm, polycarbonate, pore size 0.1 µm; Whatman)
  • Straight dissecting forceps, length 110 mm
  • Dissection microscope
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Figures

Videos

Literature Cited

Literature Cited
   Donovan, J. and Brown, P. 2006. Euthanasia. Curr. Protoc. Immunol. 73:1.8.1‐1.8.4.
   Grobstein, C. 1953. Morphogenetic interaction between embryonic mouse tissues separated by a membrane filter. Nature 172:869‐870.
   Grobstein, C. 1956. Trans‐filter induction of tubules in mouse metanephrogenic mesenchyme. Exp. Cell Res. 10:424‐440.
   Grobstein, C. 1957. Some transmission characteristics of the tubule‐inducing influence on mouse metanephrogenic mesenchyme. Exp. Cell Res. 13:575‐587.
   Kaufman, M.H. 1992. The Atlas of Mouse Development. Academic Press, London.
   Rugh, R. 1968. The Mouse: Its Reproduction and Development. Burgess Publishing Company, Minneapolis, Minn.
   Sakai, T., Larsen, M., and Yamada, K.M. 2003. Fibronectin requirement in branching morphogenesis. Nature 423:876‐881.
   Saxen, L., Salonen, J., Ekblom, P., and Nordling, S. 1983. DNA synthesis and cell generation cycle during determination and differentiation of the metanephric mesenchyme. Dev. Biol. 98:130‐138.
   Tuan, R.S. and Lo, C.W. Developmental Biology Protocols. Humana Press, Totowa, N.J.
   Wartiovaara, J., Linder, E., Ruoslahti, E., and Vaheri, A. 1974a. Distribution of fibroblast surface antigen: Association with fibrillar structures of normal cells and loss upon viral transformation. J. Exp. Med. 140:1522‐1533.
   Wartiovaara, J., Nordling, S., Lehtonen, E., and Saxen, L. 1974b. Transfilter induction of kidney tubules: Correlation with cytoplasmic penetration into nucleopore filters. J. Embryol. Exp. Morphol. 31:667‐682.
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