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ARS Home » Southeast Area » Auburn, Alabama » Aquatic Animal Health Research » Research » Publications at this Location » Publication #414954
Research Project: Integrated Research to Improve Aquatic Animal Health in Warmwater Aquaculture

Location: Aquatic Animal Health Research

Title: The single-cell transcriptome analysis of channel catfish B and T cell lines

Author
item Lange, Miles
item ALDERSEY, JOHANNA - Orise Fellow
item Beck, Benjamin
item Abernathy, Jason

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 6/23/2024
Publication Date: 6/23/2024
Citation: Lange, M.D., Aldersey, J.E., Beck, B.H., Abernathy, J.W. 2024. The single-cell transcriptome analysis of channel catfish B and T cell lines [ABSTRACT]. 14th North American Comparative Immunology Workshop, Santa Cruz, CA. June 23-26, 2024.

Interpretive Summary:

Technical Abstract: Channel catfish (Ictalurus punctatus) have been shown to be an excellent model for understanding teleost immunity, and the development of several cell lines has proved to be indispensable in many fundamental studies into the function of B and T lymphocytes. There are two B cell lines (1G8 and 3B11) developed through in vitro LPS stimulation and two T cells lines, one developed through in vitro LPS stimulation (G14D) and the other (28S.3) was spontaneously immortalized after the long-term culture of peripheral blood lymphocytes. All cell lines were cultured at 27°C, 5% CO2 in AL-4 medium consisting of equal parts of AIM V medium and L-15 medium adjusted to catfish tonicity with 10% DI H2O and supplemented with 1 mg/mL NaHCO3, Penicillin Streptomycin Glutamine solution, 50 mM 2-ME, and 4% heat-inactivated pooled catfish serum. 20,000 cells (~90% cell viability) for each were loaded onto a GEM-X chip and single-cell RNAseq libraries were prepared using the 10X Genomics Chromium platform and later sequenced (~20,000 reads/cell) on an Illumina NovaSeq X Plus. The demultiplexed samples will be aligned to the CoCo_2.0 channel catfish reference assembly and filtered using Cell Ranger (v.5.0.0). Integrated data will be analyzed in Seurat (v.5.0.1). Cluster analysis will be performed to identify any heterogeneity among the different cell lines. Differential gene expression and markers that define individual cell types will be discussed.