Biology subjects

Van de Walle, G. R.

Publications and source records attributed to Van de Walle, G. R..

2 recordsLinked to original sources

bioRxiv2021

Establishment and characterization of mammary organoids from non-traditional model organisms

Mammary organoid (MaO) models are only available for a few traditional model organisms, limiting our ability to investigate mammary gland development and cancer across the diverse taxa of mammals. For example, horses are mammals with a similar mammary anatomy and function as humans, but they have a remarkably low incidence of mammary cancer, making the development of MaOs in non-traditional model organisms attractive, particularly in comparative cancer research. This study established equine mammary organoids (EqMaOs) from mammary gland tissue fragments and evaluated parameters including diameter, budding, and growth stage in non-budding EqMaOs, in cultures with increasing concentrations of epidermal growth factor (EGF), a key growth factor implicated in mammary gland development. Our findings showed that EqMaO diameter is not influenced by EGF concentration, whereas number of EqMaOs with budding and stage in non-budding EqMaOs are positively influenced by increasing EGF concentration. EqMaOs also formed protrusions with putative functions, including organoid fusion and sensory functions. We further characterized EqMaOs by the presence of myoepithelial and luminal cells using immunohistochemistry and used the hormone prolactin to stimulate milk secretion, as illustrated by {beta}-lactoglobulin expression, in these EqMaOs. Additionally, we showed that our method to establish MaOs is widely applicable to additional non-traditional mammalian model organisms such as cat, pig, deer, rabbit, and prairie vole. Collectively, MaO models across species will be a useful tool for comparative developmental and cancer studies. Summary statementMammary organoids can be established from various mammals by embedding mammary tissue fragments into a 3D matrix, providing a high-throughput, physiologically accurate model for comparative studies centered on mammary gland development and cancer.

cell biology↗

bioRxiv2020

Single cell resolution landscape of equine peripheral blood mononuclear cells reveals diverse immune cell subtypes including T-bet+ B cells

Traditional laboratory model organisms represent a small fraction of the diversity of multicellular life, and findings in any given experimental model often do not translate to other species. Immunology research in non-traditional model organisms can be advantageous or even necessary (e.g. for host-pathogen interaction studies), but presents multiple challenges, many stemming from an incomplete understanding of potentially species-specific immune cell types, frequencies and phenotypes. Identifying and characterizing immune cells in such organisms is frequently limited by the availability of species-reactive immunophenotyping reagents for flow cytometry, and insufficient prior knowledge of cell type-defining markers. Here, we demonstrate the utility of single cell RNA sequencing (scRNA-Seq) to characterize immune cells for which traditional experimental tools are limited. Specifically, we used scRNA-Seq to comprehensively define the cellular diversity of equine peripheral blood mononuclear cells (PBMCs) from healthy horses across different breeds, ages, and sexes. We identified 30 cell type clusters partitioned into five major populations: Monocytes/Dendritic Cells, B cells, CD3+PRF1+ lymphocytes, CD3+PRF1- lymphocytes, and Basophils. Comparative analyses revealed many cell populations analogous to human PBMC, including transcriptionally heterogeneous monocytes and distinct dendritic cell subsets (cDC1, cDC2, plasmacytoid DC). Unexpectedly, we found that a majority of the equine peripheral B cell compartment is comprised of T-bet+ B cells; an immune cell subpopulation typically associated with chronic infection and inflammation in human and mouse. Taken together, our results demonstrate the potential of scRNA-Seq for cellular analyses in non-traditional model organisms, and form the basis for an immune cell atlas of horse peripheral blood.

immunology↗