Genetic Manipulation and Extended Culture of Human Germinal Center B Cells to Model Lymphomagenesis

Methods Mol Biol. 2025:2865:395-409. doi: 10.1007/978-1-0716-4188-0_17.

Abstract

The germinal center (GC) is the stage of B cell differentiation that gives rise to a majority of B cell lymphomas. Here, we present an experimental coculture system for the ex vivo expansion and genetic manipulation of human GC B cells purified from discarded tonsil tissue. This system can be used to investigate the impact of defined genetic alterations, either individually or in combination, upon the growth and survival of human GC B cells in vitro. We provide examples of genetic combinations that lead to the immortalized growth of GC B cells in vitro, and others that result in malignant transformation in immunodeficient mice, allowing the creation of genetically bespoke, synthetic, human lymphoma models.

Keywords: CRISPR; Functional genomics; Germinal center; Lymphoma; Viral transduction.

MeSH terms

  • Animals
  • B-Lymphocytes* / metabolism
  • Cell Culture Techniques / methods
  • Cell Differentiation
  • Cell Transformation, Neoplastic / genetics
  • Coculture Techniques* / methods
  • Germinal Center* / immunology
  • Germinal Center* / metabolism
  • Humans
  • Lymphoma, B-Cell / genetics
  • Lymphoma, B-Cell / pathology
  • Mice
  • Palatine Tonsil / cytology