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Isolation of Embryonic Tissues and Formation of Quail-Chicken Chimeric Organs Using The Thymus Example
Published on: February 16, 2019
Establishment of an in vitro system representing the chicken gut-associated lymphoid tissue
Noorjahan Banu Alitheen1, Susan Jane McClure, Swee Keong Yeap
1Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Insights
Researchers created in vitro cellular agglomerates mimicking the chicken bursa for B cell studies. These structures support B cell development and offer a model for studying avian diseases like Newcastle disease virus.
Area of Science:
- Immunology
- Developmental Biology
- Avian Biology
Background:
- The bursa of Fabricius is essential for B cell development in chick embryos.
- Understanding B cell differentiation requires models that replicate bursal microenvironments.
Purpose of the Study:
- To develop an in vitro system that mimics the functional aspects of the chicken bursa.
- To characterize the cellular dynamics and B cell populations within these in vitro structures.
- To assess the utility of this model for studying avian viral infections.
Main Methods:
- Co-cultivation of embryonic epithelial and lymphoid cells.
- Formation and characterization of cellular agglomerates.
- Flow cytometry to analyze lymphocyte populations (Bu-1a, IgM).
- Assessment of activation-induced deaminase expression and cell proliferation.
Main Results:
- Cellular agglomerates resembling bursal tissue formed within 48 hours.
- Agglomerates contained Bu-1a(+) lymphocytes, while emigrant cells were Bu-1a(-) and IgM(+).
- Activation-induced deaminase expression increased in both agglomerates and emigrant cells.
- Emigrating cells exhibited higher proliferation rates than the initial cell population.
Conclusions:
- In vitro generated agglomerates serve as a functional analogue of the chicken bursa.
- This model system facilitates the study of B cell differentiation and activation.
- The system shows promise for investigating avian pathogen responses, such as Newcastle disease virus infection.
Abstract:
The bursa of Fabricius is critical for B cell development and differentiation in chick embryos. This study describes the production in vitro, from dissociated cell suspensions, of cellular agglomerates with functional similarities to the chicken bursa. Co-cultivation of epithelial and lymphoid cells obtained from embryos at the appropriate developmental stage regularly led to agglomerate formation within 48 hours. These agglomerates resembled bursal tissue in having lymphoid clusters overlaid by well organized epithelium. Whereas lymphocytes within agglomerates were predominantly Bu-1a(+), a majority of those emigrating onto the supporting membrane were Bu-1a(-) and IgM(+). Both agglomerates and emigrant cells expressed activation-induced deaminase with levels increasing after 24 hours. Emigrating cells were actively proliferating at a rate in excess of both the starting cell population and the population of cells remaining in agglomerates. The potential usefulness of this system for investigating the response of bursal tissue to avian Newcastle disease virus (strain AF2240) was examined.
