Preparation of Mouse Feeder Cells for Fusion and Cloning

Steven A. Fuller1, Miyoko Takahashi1, John G.R. Hurrell1

1 Allelix Inc., Mississauga, Ontario
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Unit 11.6
DOI:  10.1002/0471142727.mb1106s01
Online Posting Date:  May, 2001
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Abstract

To maximize the yield of hybrids from fusion and cloning procedures, feeder cells are required to be cocultured with the hybrids, while hybrid cell density is low. Mouse peritoneal cells, most of which are macrophages, have been found to be convenient and effective feeder cells which are a source of soluble growth factors for hybridoma cells.

     
 
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Table of Contents

  • Reagents and Solutions
  • Commentary
     
 
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Materials

Basic Protocol 1:

  Materials
  • recipe0.34 M sucrose solution, sterile and chilled
  • Mice (any strain)
  • 70% ethanol
  • recipeHAT medium, chilled
  • Sterile phosphate‐buffered saline (PBS; appendix 22)
  • 10‐ml syringe, sterile
  • 18‐G needle, sterile
  • 50‐ml conical centrifuge tube, sterile
  • Dissecting board
  • Forceps, sterile
  • Scissors, sterile
  • 96‐well microtiter plates
  • 8% CO 2‐in‐air gas mixture
  • Humidified CO 2 incubator
  • Additional reagents and equipment for estimating cell viability by trypan blue exclusion ( support protocol, unit 11.5)
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Figures

Videos

Literature Cited

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