Duchesne Camille, Randuineau Gwénaëlle, Le Normand Laurence, Romé Véronique, Laraqui Samia, Arnaud Alexis Pierre, Boudry Gaëlle
Institut Numecan, INRAE, INSERM, Univ Rennes, Rennes, France.
CHU, Rennes, France.
Animal Model Exp Med. 2024 Dec;7(6):944-954. doi: 10.1002/ame2.12512. Epub 2024 Dec 15.
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.
Organoids were derived from jejunal and colonic ISC obtained from 1-, 7-, 28-, 36- and 180-day-old pigs and passaged three times.
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.
In conclusion, while the age of the tissue donor affected organoid growth and morphology, it did not influence the phenotype.
在减少动物实验的背景下，肠道类器官是很有前景的工具，但要常规使用这种细胞模型，需要全面表征肠道干细胞（ISC）来源对类器官表型的影响。我们的目的是评估ISC供体年龄对源自猪的肠道类器官生长、形态和细胞组成的影响。
类器官源自1日龄、7日龄、28日龄、36日龄和180日龄猪的空肠和结肠ISC，并传代三次。
我们首先通过qPCR证实，在传代两次后，超过80个与各种肠道功能相关的研究基因中，18%的基因在空肠和结肠类器官中的表达存在差异（p < 0.05）。类器官的生长和形态不仅取决于肠道位置（源自结肠的类器官数量更多且更大，比源自空肠的ISC大，p < 0.05），还取决于猪的年龄。实际上，当ISC来自幼龄仔猪时，与来自老龄猪的ISC相比，类器官与球体的比例更大（p < 0.05），原代培养期间球体更大，但传代两次后更小（p < 0.05），传代一次后类器官更小（p > 0.05）。最后，通过对主要肠道细胞类型（吸收性、肠内分泌和杯状细胞）标志物的免疫染色评估，在源自7日龄或180日龄猪的类器官之间未观察到细胞组成的差异，但注意到肠道部位来源之间存在差异。
总之，虽然组织供体的年龄影响类器官的生长和形态，但不影响其表型。
