Generation of Tumor Cells Expressing Firefly Luciferase (fLuc) to Evaluate the Effectiveness of CAR in a Murine Model

  • Marcelo de Souza Fernandes Pereira
  • Daianne Maciely Carvalho Fantacini
  • Virgínia Picanço-Castro
Part of the Methods in Molecular Biology book series (MIMB, volume 2086)


Immunotherapy has been showed as a promisor treatment, in special for hematological diseases. Chimeric antigen receptor T cells (CARs) which are showing satisfactory results in early-phase cancer clinical trials can be highlighted. However, preclinical models are critical steps prior to clinical trial. In this way, a well-established preclinical model is an important key in order to confirm the proof of principle. For this purpose, in this chapter will be pointed the methods to generate tumor cells expressing firefly Luciferase. In turn, these modified cells will be used to create a subcutaneous and a systemic murine model of Burkitt’s lymphoma in order to evaluate the effectiveness of CAR-T.

Key words

Preclinical model Subcutaneous model Systemic model Luciferase Bioluminescence imaging CAR-T 



This work was supported by São Paulo Research Foundation—FAPESP 17/09491-8, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), CTC Center for Cell-based Therapies (FAPESP 2013/08135-2), and National Institute of Science and Technology in Stem Cell and Cell Therapy (CNPq 573754-2008-0 and FAPESP 2008/578773). The authors also acknowledge financial support from Secretaria Executiva do Ministério da Saúde (SE/MS), Departamento de Economia da Saúde, Investimentos e Desenvolvimento (DESID/SE), Programa Nacional de Apoio à Atenção Oncológica (PRONON) Process 25000.189625/2016-16.


  1. 1.
    Rehemtulla A, Stegman LD, Cardozo SJ et al (2000) Rapid and quantitative assessment of cancer treatment response using in vivo bioluminescence imaging. Neoplasia 2(6):491–495CrossRefGoogle Scholar
  2. 2.
    Edinger M, Cao YA, Hornig YS et al (2002) Advancing animal models of neoplasia through in vivo bioluminescence imaging. Eur J Cancer 38(16):2128–2136CrossRefGoogle Scholar
  3. 3.
    Shimomura O, Goto T, Johnson FH (1977) Source of oxygen in the CO(2) produced in the bioluminescent oxidation of firefly luciferin. Proc Natl Acad Sci U S A 74(7):2799–2802CrossRefGoogle Scholar
  4. 4.
    Cosette J, Ben Abdelwahed R, Donnou-Triffault S et al (2016) Bioluminescence-based tumor quantification method for monitoring tumor progression and treatment effects in mouse lymphoma models. J Vis Exp 113.
  5. 5.
    Burkitt D (1958) A sarcoma involving the jaws in African children. Br J Surg 46(197):218–223CrossRefGoogle Scholar
  6. 6.
    Burkitt D, O’Conor GT (1961) Malignant lymphoma in African children. I. A clinical syndrome. Cancer 14:258–269CrossRefGoogle Scholar
  7. 7.
    Jacobson C, LaCasce A (2014) How I treat Burkitt lymphoma in adults. Blood 124(19):2913–2920CrossRefGoogle Scholar
  8. 8.
    Dalla-Favera R, Bregni M, Erikson J et al (1982) Human c-myc onc gene is located on the region of chromosome 8 that is translocated in Burkitt lymphoma cells. Proc Natl Acad Sci U S A 79(24):7824–7827CrossRefGoogle Scholar
  9. 9.
    Kelemen K, Braziel RM, Gatter K et al (2010) Immunophenotypic variations of Burkitt lymphoma. Am J Clin Pathol 134(1):127–138CrossRefGoogle Scholar
  10. 10.
    Pulvertaft JV (1964) Cytology of Burkitt’s Tumour (African Lymphoma). Lancet 1(7327):238–240CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Marcelo de Souza Fernandes Pereira
    • 1
  • Daianne Maciely Carvalho Fantacini
    • 1
  • Virgínia Picanço-Castro
    • 1
  1. 1.Center for Cell-Based Therapy CTC, Regional Blood Center of Ribeirão PretoUniversity of São PauloSão PauloBrazil

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