In-silicoandin-vitromorphometric analysis of intestinal organoids

Abstract

Organoids offer a powerful model to study cellular self-organisation, the growth of specific tissue morphologies in-vitro, and to assess potential medical therapies. However, the intrinsic mechanisms of these systems are not entirely understood yet, which can result in variability of organoids due to differences in culture conditions and basement membrane extracts used. Improving the standardisation of organoid cultures is essential for their implementation in clinical protocols. Developing tools to assess and predict the behaviour of these systems may produce a more robust and standardised biological model to perform accurate clinical studies. Here we developed an algorithm to automate crypt-like structure counting on intestinal organoids in both in-vitro and in-silico images. In addition, we modified an existing two-dimensional agent-based mathematical model of intestinal organoids to better describe the system physiology, and evaluated its ability to replicate budding structures compared to new experimental data we generated. The crypt-counting algorithm proved useful in approximating the average budding structures found in our in-vitro intestinal organoid culture images on days 3 and 7 after seeding. Our changes to the in-silico model maintain the potential to produce simulations that replicate the number of budding structures found on days 5 and 7 of in-vitro data. The present study aims to aid in quantifying key morphological structures and provide a method to compare both in-vitro and in-silico experiments. Our results could be extended later to 3D in-silico models.

MAGIC matrices: freeform bioprinting materials to support complex and reproducible organoid morphogenesis

“…By contrast, over 5 days, manually seeded organoids showed significant heterogeneity in size and maturity over the same timeframe (Figure 5B,C). Furthermore, for a given amount of time post-seeding, bioprinted organoid arrays underwent more extensive and uniform morphogenesis, exhibiting a greater number of crypts with decreased variance compared to manually seeded organoids (Figure 5D) 71 . Organoids grown in manually seeded cultures using MAGIC matrix as opposed to pure Matrigel showed no difference in crypt number (Figure 5D), indicating that the bioprinting method itself leads to this improved maturity.…”