ArticleNature protocols2021
Genetic manipulation and immortalized culture of ex vivo primary human germinal center B cells.
Article in Nature protocols, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 28 citations in OpenAlex.
- Article
- Synergistic targeting of cancer cells through simultaneous inhibition of key metabolic enzymes.Cell death and differentiation · 2025Article
- Enforced MYC expression directs a distinct transcriptional state during plasma cell differentiation.Life science alliance · 2025Article
- Audit of B-cell cancer genes.Blood advances · 2025Review
- Genetic Manipulation and Extended Culture of Human Germinal Center B Cells to Model Lymphomagenesis.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Integrative single-cell chromatin and transcriptome analysis of human plasma cell differentiation.Blood · 2024Article
- Patterns of Oncogene Coexpression at Single-Cell Resolution Influence Survival in Lymphoma.Cancer discovery · 2023Article
- Immortalized B Cells Transfected with mRNA of Antigen Fused to MITD (IBMAM): An Effective Tool for Antigen-Specific T-Cell Expansion and TCR Validation.Biomedicines · 2023Article
- Review
- Article
- Genetic Profiling in Diffuse Large B-Cell Lymphoma: The Promise and the Challenge.Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc · 2023Review
- In Vitro Diffuse Large B-Cell Lymphoma Cell Line Models as Tools to Investigate Novel Immunotherapeutic Strategies.Cancers · 2022Review
- Efficient CRISPR-Cas9-mediated mutagenesis in primary human B cells for identifying plasma cell regulators.Molecular therapy. Nucleic acids · 2022Article
- Primary B cell engineering for therapeutic research.Trends in molecular medicine · 2022Article
- SHMT2 inhibition disrupts the TCF3 transcriptional survival program in Burkitt lymphoma.Blood · 2022Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Next-generation sequencing has transformed our knowledge of the genetics of lymphoid malignancies. However, limited experimental systems are available to model the functional effects of these genetic changes and their implications for therapy. The majority of mature B-cell malignancies arise from the germinal center (GC) stage of B-cell differentiation. Here we describe a detailed protocol for the purification and ex vivo expansion of primary, nonmalignant human GC B cells. We present methodology for the high-efficiency transduction of these cells to enable combinatorial expression of putative oncogenes. We also describe alternative approaches for CRISPR-Cas9-mediated deletion of putative tumor suppressors. Mimicking genetic changes commonly found in lymphoid malignancies leads to immortalized growth in vitro, while engraftment into immunodeficient mice generates genetically customized, synthetic models of human lymphoma. The protocol is simple and inexpensive and can be implemented in any laboratory with access to standard cell culture and animal facilities. It can be easily scaled up to enable high-throughput screening and thus provides a versatile platform for the functional interrogation of lymphoma genomic data.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.