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Survival Surgery: Removal of the Spleen or Thymus

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

1Food and Drug Administration, Bethesda, Maryland
2University 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

  • Unit Introduction
  • Basic Protocol: Splenectomy
  • Alternate Protocol: Hemisplenectomy
  • Basic Protocol: Neonatal Thymectomy
  • Basic Protocol: Adult Thymectomy
  • Support Protocol: Construction of Cannula for Suction Thymectomy
  • Commentary
  • Bibliography
  • Figures
     
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Materials

Basic Protocol: 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)

Basic Protocol: Neonatal Thymectomy

 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 support 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: Adult 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 (support 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)
Looking for materials?  Suppliers Guide
     
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Figures

  • Figure 1.10.1
    Surgical removal of the spleen. Through the small incisions in the skin, abdominal musculature, and peritoneum, the spleen is exteriorized. The splenic artery is ligated and the spleen removed.

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  • Figure 1.10.2
    Neonatal thymectomy. (A) Position of anesthetized newborn mouse secured to the operating board with rubber bands. Thymus and surrounding structures following sternal incision (blowup). (B) Position of instruments to remove thymus from neonate is indicated.

    View Image
  • Figure 1.10.3
    Adult thymectomy. (A) Position of the adult mouse secured to the operating board with rubber bands. (B) Location of the sternal notch. The incision is made in the midline from the sternal notch to the second or third rib (blowup).

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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 . 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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