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Increased initiation and growth of tumor cell lines, cancer stem cells and biopsy material in mice using basement membrane matrix protein (Cultrex or Matrigel) co-injection

Nature Protocols volume 7, pages 1138–1144 (2012)Cite this article

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Abstract

This protocol requires 2–4 h and presents a method for injecting tumor cells, cancer stem cells or dispersed biopsy material into subcutaneous or orthotopic locations within recipient mice. The tumor cells or biopsy are mixed with basement membrane matrix proteins (CultrexBME or Matrigel) at 4 °C and then injected into recipient animals at preferred anatomical sites. Tumor cells can also be co-injected with additional cell types, such as fibroblasts, stromal cells, endothelial cells and so on. Details are given on appropriate cell numbers, handling and concentration of the basement membrane proteins, recipient animals, injection location and techniques. This procedure enables the growth of tumors from cells or biopsy material (tumor graft) with greater efficiency of take and growth, and with retention of the primary tumor phenotype based on histology. Co-injection with additional cell types provides more physiological models of human cancers for use in drug screening and studying cancer biology.

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Figure 1: Macroscopic appearance and vessel infiltration of prostate tumors 9 d after s.c. injection.
Figure 2: Appearance of mammary fat pad breast tumors grown in the presence and absence of CultrexBME/Matrigel.

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Acknowledgements

The authors want to acknowledge the pioneering work and leading role of G.R. Martin in this area and are grateful for his support and contribution.

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Author notes

  1. hkleinman@dir.nidcr.nih.gov

Authors and Affiliations

  1. Department of Pathology, Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan, USA

    R Daniel Bonfil

  2. Trevigen, Inc., Gaithersburg, Maryland, USA

    Gabriel Benton

  3. Department of Biochemistry and Molecular Biology, The George Washington University Medical Center, Washington, District of Columbia, USA

    Irina Aranoutova & Hynda K Kleinman

  4. Department of Urology, Wayne State University School of Medicine, Karmanos Cancer Institute, Detroit, Michigan, USA

    R Daniel Bonfil

  5. Rafael Fridman

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  1. Rafael Fridman
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  4. Hynda K Kleinman
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Contributions

R.F. and R.D.B. performed the experiments and optimized certain aspects of the procedure. H.K.K. performed the experiments. G.B. and I.A. optimized preparation of the basement membrane proteins. H.K.K. wrote the manuscript with assistance from R.F., R.D.B., G.B. and I.A.

Corresponding authors

Correspondence to Rafael Fridman or Hynda K Kleinman.

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Competing interests

R.F. and R.D.B. have no financial interests. H.K.K., G.B. and I.A. declare competing financial interests. G.B. and I.A are employees of Trevigen Inc., and H.K.K. is a consultant for Trevigen Inc. which is one of the companies commercializing the basement membrane extract (CultrexBME/Matrigel).

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Fridman, R., Benton, G., Aranoutova, I. et al. Increased initiation and growth of tumor cell lines, cancer stem cells and biopsy material in mice using basement membrane matrix protein (Cultrex or Matrigel) co-injection. Nat Protoc 7, 1138–1144 (2012). https://doi.org/10.1038/nprot.2012.053

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