Atili et al 2019

Calcium-induced differentiation in normal human colonoid cultures: Cell-cell / cell- matrix adhesion, barrier formation and tissue integrity

D Attili, SD McClintock, AH Rizvi, S Pandya, H Rehman, DM Nadeem, A Richter, D Thomas, MK Dame, DK Turgeon, J Varani, MN Aslam
Published: PLoS ONE 14(4): 2019

Background:

Progression from initial polyp formation to more serious disease is difficult to study experimentally since colonic polyps are removed upon detection. Colonoid culture technology provides a way to study human colon polyp responses to potentially useful compounds under ex-vivo conditions.
The goal of the study was to assess calcium alone against Aquamin for capacity to affect growth and differentiation in colonoid cultures derived from histologically-normal human colon tissue (n=5).

Methods: 

Colonoid cultures were maintained in a low-calcium (0.25 mM) medium or in medium supplemented with an amount of calcium (1.5–3.0 mM), either from calcium alone or Aquamin for two weeks.. Changes in growth, morphological features and protein expression profiles were then assessed using a combination of phase-contrast, scanning electron & transmission-electron microscopy, histology and immunohistology, & proteomic assessment.

Conclusions : 

Here we show in colonoids derived from histologically-normal tissue that extensive differentiation occurred in the absence of intervention, and that treatment with either calcium alone or Aquamin had only a modest additional effect in the gross and histological appearance of normal tissue-derived colonoids. Additionally, there was a strong induction of several proteins that contribute to cellular adhesive functions, barrier formation and tissue integrity.

This is in contrast to previous studies in adenoma colonoids where Aquamin suppressed growth and induced features of differentiation not seen with calcium alone at a comparable level (McClintock et al., 2018).

Figure:

Colonoid appearance in culture: Histological features. At the end of the incubation period, colonoids were examined by light microscopy

(lower panels) Cell surface components were examined between colonoids.

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