Survival Surgery: Removal of the Spleen or Thymus

J.P. Reeves1, P.A. Reeves1, L. Thomas Chin2

1 Food and Drug Administration, Bethesda, Maryland, 2 University of Massachusetts, Worcester, Massachusetts
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 1.10
DOI:  10.1002/0471142735.im0110s02
Online Posting Date:  May, 2001
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Abstract

Although for most experiments lymphoid tissue is removed from the freshly sacrificed animal, there are several circumstances which require the surgical removal, under anesthesia, of either the thymus or spleen, and the recovery of the animal for further investigation. This unit describes survival surgery for removal of the adult spleen as well as the adult and neonatal thymus of the mouse.

     
 
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Table of Contents

  • Basic Protocol 1: Splenectomy
  • Alternate Protocol 1: Hemisplenectomy
  • Basic Protocol 2: Neonatal Thymectomy
  • Basic Protocol 3: Adult Thymectomy
  • Support Protocol 1: Construction of Cannula for Suction Thymectomy
  • Commentary
  • Figures
     
 
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Materials

Basic Protocol 1: Splenectomy

  Materials
  • Mouse (>3 weeks old)
  • 1 mg/ml pentobarbital
  • 70% ethanol
  • Iris scissors (straight or curved)
  • Iris forceps (10 cm long with curved ends)
  • 4‐0 sutures
  • Sterile 10 × 10–cm gauze sections
  • Additional reagents and equipment for anesthesia (unit 1.4) and intraperitoneal injection (unit 1.6)

Alternate Protocol 1: Hemisplenectomy

  Materials
  • Mouse (0 to 3 days old)
  • Saline or phosphate‐buffered saline (PBS; appendix 2A)
  • Sterile 5 × 5–cm gauze sections
  • Incandescent desk lamp
  • Dissecting board (wood or styrofoam)
  • Dissecting microscope
  • Iris scissors (angled with extra‐fine tips; Roboz #RS‐5650, ‐5658 or equivalent)
  • Forceps with nonserrated ends (curved with fine tips; Roboz #RS‐5045, ‐5047, or ‐5057 or equivalent)
  • Cannula (see protocol 5support protocol)
  • Hemostats or forceps
  • 6‐0 suture
  • Cotton‐tipped applicators, soaked in saline or PBS
  • Topical flexible collodion (U.S.P.)
  • 60‐ or 100‐mm Petri plate, sterile

Basic Protocol 2: Neonatal Thymectomy

  Materials
  • Mouse (>3 weeks old)
  • 70% ethanol
  • Dissecting board (∼7 × 10–cm)
  • Elastic bands
  • Iris scissors (10‐cm straight)
  • Iris forceps (2 pair of 10‐cm half‐curved)
  • Sterile 5 × 5–cm gauze sections
  • Suction cannula ( protocol 5support protocol)
  • Rubber tubing, 6‐mm diameter
  • Clay Adams autoclip applier (#7630‐000‐900) and 9‐mm wound clips (#7631)
  • Incandescent desk lamp
  • Additional reagents and equipment for anesthesia (unit 1.4) and intraperitoneal injection (unit 1.6)
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Figures

Videos

Literature Cited

Literature Cited
   Bandeira, A., Itohara, S., Bonneville, M., Burlen‐Defranoux, O., Mota‐Santos, T., Coutinho, A., and Tonegawa, S. 1991. Extrathymic origin of intestinal epithelial lymphocytes bearing T cell antigen receptor γδ. Proc. Natl. Acad. Sci. U.S.A. 88:43‐47.
   Miller, J.F.A.P. 1964. The thymus and the development of immunologic responsiveness. Science 144:1544‐1547.
   Miller, J.F.A.P. 1965. Effects of thymectomy in adult mice on immunological responsiveness. Nature 208:1337‐1339.
   Pantelouris, E.M. 1968. Absence of thymus in a mouse mutant. Nature 217:370‐371.
   Qin, S., Cobbold, S., Benjamin, R., Waldmann, H. 1989. Induction of classical transplantation tolerance in the adult. J. Exp. Med. 169:779.
   Sprent, J. 1977. Migration and lifespan of lymphocytes. In B and T Cells in Immune Recognition, pp. 59‐82. John Wiley & Sons, London.
   Webb, S.R., Morris, C., Sprent, J.S. 1990. Extrathymic tolerance of mature T cells: Clonal elimination as a consequence of immunity. Cell 63:1249‐1256.
Key Reference
   Sjokin, K., Dalmasso, A.P., Smith, J.M., and Martinez, C. 1963. Thymectomy in newborn and adult mice. Transplantation 1:521‐525.
  Classic description of method of neonatal and adult thymectomy.
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