Morphological alterations of cultured human colorectal matched tumour and healthy organoids

Abstract

Organoids have extensive applications in many fields ranging from modelling human development and disease, personalised medicine, drug screening, etc. Moreover, in the last few years, several studies have evaluated the capacity of organoids as transplantation sources for therapeutic approaches and regenerative medicine. Nevertheless, depending on the origin of the cells and anatomical complications, an organoid transplant may make tissue regeneration difficult. However, some essential aspects of organoids including the morphological alterations and the growth pattern of the matched tumour and their healthy derived organoids have received less attention. Therefore, the current work focused on culturing matched healthy and tumour organoids from the same patient with colorectal cancer (CRC) and assessed their timed growth and structural differences on a daily basis. The healthy organoids underwent proliferation and branching morphogenesis, while the tumour organoids did not follow the same pattern, and the majority of them developed cystic structures instead. However, the number and size of tumour organoids were different from one patient to another. The differential morphological changes of the healthy versus human colonic tumour organoids likely linked to distinct molecular and cellular events during each day. Thus, while their specific structural features provide valuable in vitro models to study various aspects of human intestinal/colon tissue homeostasis and CRC which avoid or replace the use of animals in research, this model may also hold a great promise for the transplantation and regenerative medicine applications.

Repurposing Antibacterial AM404 As a Potential Anticancer Drug for Targeting Colorectal Cancer Stem-Like Cells

“…Thus, several established stem cell markers, CRC prognostic factors, and differentiation markers have also revealed the potential of AM404 in targeting stem-like cells. As a first step towards mimicking the patient tumour tissues, we used CRC patients' derived organoids [42,43], to study specific cell-type response to drugs [44]. Organoids were cultured and allowed to start budding for 5-6 days prior to treating them with AM404.…”

“…Tumour and adjacent healthy tissue samples of 17 CRC patients were collected [43] from Nottingham Health Sciences Biobank (NHSB), Queens Medical Centre, University of Nottingham. Ethical approval and research and development approval including written informed consent were obtained by Nottingham Health Science Biobank (NHSB), Histopathology Department, School of Medicine, University of Nottingham, Nottingham NG7 2UH.…”

“…CRC patient-derived organoids were cultured as previously established and characterized in our lab [42,43]. Colonic crypts were made to release from intestinal epithelium and were re-suspended in Matrigel in the presence of complete organoid medium [43].…”

“…CRC patient-derived organoids were cultured as previously established and characterized in our lab [42,43]. Colonic crypts were made to release from intestinal epithelium and were re-suspended in Matrigel in the presence of complete organoid medium [43]. Once the organoids started to grow (usually day 4-5), they were treated with drug and/or complete organoid medium as vehicle control for their morphology, growth evaluation and live/dead staining.…”

Replication Study: Wnt activity defines colon cancer stem cells and is regulated by the microenvironment

“…Furthermore, recent studies have continued to examine the role of the microenvironment and cancer stemness. Niche factor requirements in colorectal tumors were found to decrease during tumorigenesis (Fujii et al, 2016; Kashfi et al, 2018). While a new modeling approach suggested stem cell functionality during colorectal tumor expansion was defined by secreted factors from CAFs rather than cell-intrinsic properties (Flanagan et al, 2018; Lenos et al, 2018).…”