Initial pig developmental stage influences intestinal organoid growth but not cellular composition

Q1 Health Professions

Animal models and experimental medicine Pub Date : 2024-12-15 DOI:10.1002/ame2.12512

Camille Duchesne, Gwénaëlle Randuineau, Laurence Le Normand, Véronique Romé, Samia Laraqui, Alexis Pierre Arnaud, Gaëlle Boudry

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Abstract

Background

Intestinal organoids are promising tools in the context of animal experiment reduction but a thorough characterization of the impact of the origin of intestinal stem cells (ISC) on organoid phenotype is needed to routinely use this cellular model. Our objective was to evaluate the effect of ISC donor age on the growth, morphology and cellular composition of intestinal organoids derived from pig.

Methods

Organoids were derived from jejunal and colonic ISC obtained from 1-, 7-, 28-, 36- and 180-day-old pigs and passaged three times.

Results

We first confirmed by qPCR that the expression of 18% of the >80 studied genes related to various intestinal functions differed between jejunal and colonic organoids after two passages (p < 0.05). Growth and morphology of organoids depended on intestinal location (greater number and larger organoids derived from colonic than jejunal ISC, p < 0.05) but also pig age. Indeed, when ISC were derived from young piglets, the ratio of organoids to spheroids was greater (p < 0.05), spheroids were larger during the primary culture but smaller after two passages (p < 0.05) and organoids were smaller after one passage (p > 0.05) compared to ISC from older pigs. Finally, no difference in cellular composition, evaluated by immunostaining of markers of the major intestinal cell types (absorptive, enteroendocrine and goblet cells) was observed between organoids originating from 7- or 180-day-old pigs, but differences between intestinal site origins were noticed.

Conclusion

In conclusion, while the age of the tissue donor affected organoid growth and morphology, it did not influence the phenotype.

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猪的初始发育阶段会影响肠器官的生长，但不会影响细胞的组成。

背景：在动物实验还原方面，肠道类器官是一种很有前景的工具，但要常规使用这种细胞模型，还需要彻底确定肠道干细胞（ISC）来源对类器官表型的影响。我们的目的是评估肠干细胞供体年龄对猪肠道类器官生长、形态和细胞组成的影响：方法：从 1、7、28、36 和 180 日龄猪的空肠和结肠 ISC 中提取器官组织，并进行三次传代：结果：我们首先通过 qPCR 证实，与来自老龄猪的 ISC 相比，空肠和结肠有机体经过两次传代后，与各种肠道功能相关的超过 80 个研究基因中有 18% 的表达存在差异（p 0.05）。最后，通过对主要肠道细胞类型（吸收细胞、肠内分泌细胞和鹅口疮细胞）的标记物进行免疫染色，观察到来自 7 天龄或 180 天龄猪的有机体在细胞组成方面没有差异，但注意到不同肠道部位的有机体之间存在差异：总之，组织供体的年龄会影响类器官的生长和形态，但不会影响表型。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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