人们对建立替代方法来代替常规动物试验以评估化学品的发育和生殖毒性(DART)越来越感兴趣。类胃病是多能干细胞的3D聚集体,自发地表现出类似于原肠胚形成的轴向伸长形态发生。它们已被用作体外胚胎发生模型,用于发育和毒理学研究。这里,根据ICHS5(R3)指南,对基于小鼠gastruoid的测定进行了DART评估,提供了啮齿动物体内各种参考药物的血浆浓度数据,特别是NOAEL和LOAEL的Cmax和AUC。首先,根据形态学影响确定参比药物及其已知代谢物对细胞发育的不利影响浓度,即生长减少或伸长异常。然后,将从gastruloid测定获得的NOAEL至LOAEL浓度范围与啮齿动物中的NOAEL浓度范围进行比较,以检查每种化学物质的体外和体内测定之间的敏感性相似性.对于24种参考药物中的18种,在啮齿动物中同时具有NOAEL和LOAEL信息,在8倍浓度范围内,gastruloid试验的灵敏度与体内试验相当.对于8种其他参考药物中只有啮齿动物NOAEL或LOAEL信息的7种,gastruloid分析与体内数据一致。总之,这些结果支持gastruoid测定的有效性,可以用作DART评估的非动物替代方法。
There is growing interest in establishing alternative methods in place of conventional animal tests to assess the developmental and reproductive toxicity (DART) of chemicals. Gastruloids are 3D aggregates of pluripotent stem cells that spontaneously exhibit axial elongation morphogenesis similar to
gastrulation. They have been explored as in vitro embryogenesis models for developmental and toxicological studies. Here, a mouse gastruloid-based assay was validated for DART assessment in accordance with the ICH S5(R3)
guideline, which provides the plasma concentration data of various reference drugs in rodents, specifically Cmax and AUC for NOAEL and LOAEL. First, adverse effect concentrations of the reference drugs and their known metabolites on gastruloid development were determined based on morphological impact, namely reduced growth or aberrant elongation. Then, the NOAEL to LOAEL concentration range obtained from the gastruloid assay was compared with that in rodents to examine similarities in sensitivity between the in vitro and in vivo assays for each chemical. For 18 out of the 24 reference drugs that have both NOAEL and LOAEL information in rodents, the sensitivity of the gastruloid assay was comparable to the in vivo assay within an 8-fold concentration margin. For 7 out of the 8 additional reference drugs that have only NOAEL or LOAEL information in rodents, the gastruloid assay was in line with the in vivo data. Altogether, these results support the effectiveness of the gastruloid assay, which may be exploited as a non-animal alternative method for DART assessment.