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T Cell Enrichment By Nonadherence to Nylon

Karen S. Hathcock1

1National Cancer Institute, Bethesda, Maryland

Unit Number: 
Unit 3.2
DOI: 
10.1002/0471142735.im0302s30
Online Posting Date: 
May, 2001
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Abstract

Differential adherence properties of T cells, B cells, and accessory cells such as macrophages or other APC (antigen-presenting cells) can be employed to enrich for T cells. While the basis for this adherence is not understood, either nylon wool or Sephadex G-10 can be used as column matrices to which B cells and accessory cells will more readily adhere than T cells. Such procedures offer a convenient, although imprecise, means of enriching T cells through removal of accessory and B cells; use of nylon wool is preferred if both of the latter subsets are to be removed, while Sephadex is used when the goal is primarily to remove accessory cells.

     
 
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Table of Contents

  • Unit Introduction
  • Basic Protocol: T Cell Enrichment Using Nylon Wool Columns
  • Support Protocol: Preparation of Sterilized Nylon Wool Column
  • Commentary
  • Literature Cited
  • Tables
     
 
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Materials

Basic Protocol: T Cell Enrichment Using Nylon Wool Columns

 Materials
  • Complete RPMI-5 medium (appendix 2A)
  • 50-ml conical tube
  • Sterilized nylon wool column (see Support Protocol and Table 3.2.1)
  • 3-way disposable stopcock
  • 19-G and 23-G needles
  • Sorvall H-1000B rotor (or equivalent)
  • Additional reagents and equipment for preparing single-cell suspensions with removal of red blood cells (unit 3.1) and for cell viability test using trypan blue exclusion (appendix 3A)
     
    Table 3.2.1 Nylon Wool Column Specifications for T Cell Enrichment

    Column size (ml)Nylon wool (g)Loading vol. (ml)Cell capacity (108)Collection vol. (ml)

    120.8-1.021.515-18
    201.6-2.043.015-18

Support Protocol: Preparation of Sterilized Nylon Wool Column

 Additional Materials
  • 1% HCl
  • Scrubbed nylon fiber/wool (Table 3.2.1; Polysciences)
  • Asbestos gloves
  • Canine grooming brushes (2)
  • 12- or 20-ml disposable syringe (see basic protocol)
     
 
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Literature Cited

Literature Cited
    Julius, M.H., Simpson, E., and Herzenberg, L.A. 1973. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur. J. Immunol. 3:645-649.
     
 
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