Mouse SaI Sarcoma Tumor Model

Suzanne Ostrand‐Rosenberg1

1 University of Maryland Baltimore County, Baltimore, Maryland
Publication Name:  Current Protocols in Immunology
Unit Number:  Unit 20.3
DOI:  10.1002/0471142735.im2003s39
Online Posting Date:  May, 2001
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Abstract

SaI is an A/J‐derived (H‐2KkDd) transplantable mouse fibrosarcoma that produces a solid tumor when inoculated subcutaneously, intradermally, or intramuscularly, and an ascites tumor when inoculated intraperitoneally into syngeneic mice. This unit describes the establishment of primary solid SaI/N tumor, the establishment of SaI ascites tumor, and the removal for sampling of in vivo‐passaged SaI ascites tumor cells in mice.

     
 
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Table of Contents

  • Basic Protocol 1: Establishing SaI Solid Tumor in Mice
  • Basic Protocol 2: Establishing SaI Ascites Tumor in Mice
  • Support Protocol 1: Removal of In Vivo–Passaged Ascites Cells
  • Commentary
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1: Establishing SaI Solid Tumor in Mice

  Materials
  • SaI/N tumor cells (ATTC; also available from author)
  • Tissue culture medium: MEM, DMEM, or IMDM with and without 10% FBS ( appendix 2A) and 1× antibiotic‐antimycotic (Life Technologies)
  • 6‐ to 8‐week‐old, syngeneic A/J mice, male or female (The Jackson Laboratory)
  • 1‐ml tuberculin syringes with 25‐G needles
  • Additional reagents and equipment for establishing tumors (as for 4T1 mammary carcinoma; unit 20.2)

Basic Protocol 2: Establishing SaI Ascites Tumor in Mice

  Materials
  • SaI tumor cells (ATCC; also available from author)
  • Tissue culture medium: MEM, DMEM, or IMDM with and without 10% FBS ( appendix 2A) and 1× antibiotic‐antimycotic (Life Technologies)
  • 6‐ to 8‐week‐old, syngeneic A/J mice, male or female (The Jackson Laboratory)
  • 15‐ml conical tubes, sterile
  • Benctop centrifuge
  • 3‐ml syringes with 25‐G needles
  • Additional reagents and equipment for intraperitoneal injection of mice (unit 1.60)

Support Protocol 1: Removal of In Vivo–Passaged Ascites Cells

  Materials
  • A/J mice carrying SaI ascites tumor (see protocol 2)
  • 70% ethanol in a spray bottle
  • PBS, sterile ( appendix 2A)
  • Dissecting board
  • Dissecting instruments: scissors and forceps (sacrificed animals)
  • 15‐ml conical test tubes, sterile
  • Low‐speed centrifuge
  • 20‐ and 25‐G needles
  • Tabletop centrifuge (e.g., IEC Clinical)
  • Additional reagents and equipment for mouse euthanasia by CO 2 asphyxiation (unit 1.8) or anesthesia with methoxyflurane (unit 1.4)
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Figures

Videos

Literature Cited

Literature Cited
   Armstrong, T., Clements, V., Martin, B., Ting, J., and Ostrand‐Rosenberg, S. 1997. Major histocompatibility complex class II‐transfected tumor cells present endogenous antigen and are potent inducers of tumor‐specific immunity. Proc. Natl. Acad. Sci. U.S.A. 94:6886‐6891.
   Armstrong, T., Clements, V., and Ostrand‐Rosenberg, S. 1998. MHC class II‐transfected tumor cells directly present antigen to tumor‐specific CD4+ T lymphocytes. J. Immunol. 160:661‐666.
   Baskar, S., Glimcher, L., Nabavi, N., Jones, R., and Ostrand‐Rosenberg, S. 1995. Major histocompatibility complex class II+B7‐1+ tumor cells are potent vaccines for stimulating tumor rejection in tumor‐bearing mice. J. Exp. Med. 181:619‐629.
   Baskar, S., Clements, V., Glimcher, L., Nabavi, N., and Ostrand‐Rosenberg, S. 1996. Rejection of MHC class II‐transfected tumor cells requires induction of tumor‐encoded B7‐1 and/or B7‐2 costimulatory molecules. J. Immunol. 156:3821‐3827.
   Fox, J., Cohen, B., and Loew, F. 1984. Laboratory Animal Medicine, p. 721. Academic Press.
   Leach, D., Krummel, M., and Allison, J. 1996. Enhancement of antitumor immunity by CTLA‐4 blockade. Science 271:1734‐1736.
   Ostrand‐Rosenberg, S., Thakur, A., and Clements, V. 1990. Rejection of mouse sarcoma cells after transfection of MHC class II genes. J. Immunol. 144:4068‐4071.
   Qi, L. and Ostrand‐Rosenberg, S. 2000. MHC class II‐restricted ER‐localized endogenous antigen traffics via the endocytic pathway and is unaffected by H‐2M expression in cell‐based cancer vaccines. Traffic. 1:152‐160.
Key References
   Baskar et al., 1995. See above.
  Describes the growth of SaI/N tumors and therapy with SaI transfectants.
   Baskar et al., 1996. See above.
  Describes the removal and testing of in vivo passaged SaI ascites cells.
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