The migration and fusion events related to ROCK activity strongly influence the morphology of chicken embryo intestinal organoids

Małgorzata Pierzchalska , Małgorzata Panek , Maja Grabacka

Abstract

The method of organoid culture has become a tool widely used in gastrointestinal research, but so far, the migration of organoids derived from gut epithelium and formed in 3D Matrigel matrix has not been reported and studied. The intestinal epithelial tissue derived from 19-day-old chicken embryo was cultured in Matrigel and the dynamic properties of the forming organoids were analyzed by time-lapse image analysis. It was observed that about one in ten organoids actively moved through the matrix, at a speed of 10–20 μm/h. Moreover, rotation was observed in the majority of organoids that did not migrate long distances. The fusion events took place between organoids, which collided during the movement or growth. In our previous paper, we showed that the presence of Toll-like receptor 4 ligand, Escherichia coli lipopolysaccharide (LPS, 1 μg/ml), increased the mean organoid diameter. Here, we confirm this result and demonstrate that the Rho-associated protein kinase (ROCK) inhibitor Y-27632 (10 μM) did not completely abolish organoid migration, but prevented the fusion events, in both LPS-treated and untreated cultures. In consequence, in the presence of Y-27632, the differences between cultures incubated with and without LPS were not visible. We conclude that migration and fusion of organoids may influence their morphology and suggest that these phenomena should be taken into account during the design of experimental settings.
Author Małgorzata Pierzchalska (FoFT / DoFB)
Małgorzata Pierzchalska,,
- Department of Food Biotechnology
, Małgorzata Panek (FoFT / DoFB)
Małgorzata Panek,,
- Department of Food Biotechnology
, Maja Grabacka (FoFT / DoFB)
Maja Grabacka,,
- Department of Food Biotechnology
Journal seriesProtoplasma, ISSN 0033-183X, e-ISSN 1615-6102, (N/A 70 pkt)
Issue year2019
Vol256
No2
Pages575-581
Publication size in sheets0.5
Keywords in EnglishIntestinal organoids, Collective cell movement, Organoid fusion and rotation, Rho-associated protein kinase, Lipopolysaccharide, Chicken
ASJC Classification2700 General Medicine; 1110 Plant Science; 1307 Cell Biology
DOIDOI:10.1007/s00709-018-1312-3
URL https://link.springer.com/content/pdf/10.1007%2Fs00709-018-1312-3.pdf
Internal identifierWTŻ/2019/24
Languageen angielski
LicenseJournal (articles only); author's original; Uznanie Autorstwa (CC-BY); after publication
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The migration and fusion events related to ROCK activity strongly influence the morphology of chicken embryo intestinal organoids of 06-02-2020
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Score (nominal)70
Score sourcejournalList
Publication indicators WoS Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 0.962; WoS Impact Factor: 2018 = 2.633 (2) - 2018=2.583 (5)
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FinansowanieThe financial support for this study was provided by the Polish National Science Center (grant no. 2013/09/B/NZ9/00285 to M. Pierzchalska).
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* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
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