Southampton Cellular Research And Tissue Engineering Systems

Our research

Our proof-of-concept experiments use different culture systems to demonstrate their potential. We investigate new targets, proof-of-concept drug development and host-pathogen interactions.

We are developing and characterising highly flexible cell culture platforms that can be used to investigate a wide range of respiratory and infectious conditions. We explore if laboratory lung infections in 3D models better recapitulate in vivo events in comparison to traditional 2D systems.

  • Microscopic grayscale image of a collection of irregularly shaped cells in a culture. Some cells resembling elongated ovals, while others appear more spindle-like.
    Submerged
  • Microscope image of a closely packed collection of cellular structures, appearing in shades of gray and pink, exhibiting varying stages of cell development or differentiation.
    Differentiation d0
  • Microscope image of a bright and irregular cloud-shaped organoid, with a radiant glow and diffused edges, centered in a dark, diamond-shaped background with light edges.
    Aggregation
  • Microscopic image of a cluster of cells tightly packed together with pronounced cell membranes and granular cytoplasm at 20x magnification. There is variation in the clarity and colour intensity among the cells, ranging from lighter, more transparent regions to darker, more opaque areas.
    Differentiation d12

Organoids are complex, self-organizing microtissues grown embedded within 3D extracellular matrix. They can contain multiple differentiated cell types, exhibit cellular polarization, and often possess a central lumen and other in vivo–like architectural features.

Organoids are capable of long-term expansion in culture while remaining phenotypically and genetically stable. Primary patient-derived organoids have been described for various tissues, both healthy and cancerous.

Organoids are invaluable preclinical models for studying cancer and offer many advantages over existing human or non-human animal cancer models.

Ex vivo nasal or bronchial samples, obtained by brushing or biopsy, in addition to human pluripotent stem cells and immortalised epithelial cell lines, provide human airway epithelial cells for in vitro two-dimensional (2D) culture models as submerged monocultures. Air–liquid interface (ALI) cultures are three-dimensional (3D) epithelial culture systems where basal cells are able to differentiate in a pseudo-stratified epithelium resembling the area of the lung they originate from.

Videos of bronchial organoids

The model can be enriched in co-culture systems where the epithelium differentiates in the presence of pathogens and immune cells such as macrophages. Further complexity is achieved by the addition of fibroblasts and mesenchymal cells to mimic epithelial–mesenchymal cross-talk. To recapitulate the extracellular matrix (ECM) present in the lung interstitium, natural or synthetic macromolecules can be added to the cultures to promote the self-assembly of a mature 3D ECM.

Similarly, 3D cultures made out of cells encapsulated in macromolecule-enriched media support the growth of organoids. To investigate pathophysiological questions and in particular inflammatory processes, one example is tuberculosis granulomas, which can be developed using collagen–alginate microspheres generated by bioelectrospray methodology incorporating Mycobacterium tuberculosis-infected peripheral blood mononuclear cells.

Among other cell types, alveolar cells can be isolated from healthy surgically resected lung tissue that together with lung fibroblasts support 3D cultures mimicking the lung parenchyma. Availability of lung tissue may be exploited in ex vivo lung tissue models using small lung sections preserving its complex native architecture. Whole lung or tissue derived from pneumonectomies allows the development of human ex vivo whole-lung perfusion models or provides precision-cut lung slices (PCLSs), both preserving native structural features. Human embryonic/fetal lung explant models support lung development studies. Fetal tissue is collected 7–21 weeks post-conception, and the mixed cell population is cultured as monolayers, organoids and other 3D cultures.

In vitro and ex vitro human lung models

An infographic detailing multiple in vitro and ex vivo human lung models. At the centre of the image lies a stylized figure of a human respiratory system, showcasing the left and right lung lobes, a bronchial tree and an alveolar branch. Surrounding this central figure are various illustrations and labels that categorize different scientific models and tools used to study lung physiology. These are grouped into sections with a small diagram or illustration that provides a visual representation of the concept, such as tissue samples, cells, or laboratory equipment. The bottom left of the image includes a legend with key for various cell types, indicated by small icons.
Humbert MV, Spalluto CM, Bell J, Blume C, Conforti F, Davies ER, Dean LSN, Elkington P, Haitchi HM, Jackson C, Jones MG, Loxham M, Lucas JS, Morgan H, Polak M, Staples KJ, Swindle EJ, Tezera L, Watson A, Wilkinson TMA. Eur Respir J. 2022

Miniaturised models such as lung-on-a-chip combine microfluidics, engineering and cell biology to recreate certain aspects of organ physiology in vitro, and are a promising alternative to conventional models. Specific devices allow mimicking normal breathing patterns and enable microscopic tissue evaluation post-stretching. Other micro-physiological systems permit relevant cells of the respiratory tree to be cultured within a biopolymer or tissue-derived matrix within microfluidic devices incorporating flow. These systems can have integrated sensors to maintain cultures and monitor responses.

PNEC: pulmonary neuroendocrine cell; Th1: T-helper 1; Treg: T regulatory; NK: natural killer; DC: dendritic cell. Figure partially created with BioRender.com.