FRAME Alternatives Laboratories, School of Biomedical Sciences, University of Nottingham, Nottingham NG7 2UH, United Kingdom.
Reprod Toxicol. 2009 Dec;28(4):503-10. doi: 10.1016/j.reprotox.2009.07.003. Epub 2009 Jul 29.
The chick micromass culture system has advantages over the validated rat system - ready availability and non-culling of the donor parent - but needs to give comparable results. This study confirmed comparability and the ability to extend the system to cover cardiac effects. It was also compared with the validated embryonic stem cell cardiomyocyte model. A teratogen and paired non-teratogen with known in vivo effects were used. Differential effects were measured via changes in cell protein content, cell viability (resazurin reduction and neutral red uptake), and cell contractility. Results showed that teratogens [L-ethionine, 5-fluorouracil and sulphisoxazole] could be distinguished from non-teratogens [DL-methionine, 6-methyluracil and sulphanilamide respectively]. Dichloroacetone and dichloropropanol affected embryonic stem cells but not the micromass; dichloropropanol had a greater effect than dichloroacetone. This approach revealed differential effects on contractility independent of effects on activity/viability, whilst the total cell protein remained unchanged. We suggest that pre-validation of this system should be examined.
鸡微量组织培养系统优于经过验证的大鼠系统——供体来源易得且无需淘汰——但需要提供可比的结果。本研究证实了其可比性,并能够将该系统扩展到涵盖心脏效应。它还与经过验证的胚胎干细胞心肌细胞模型进行了比较。使用了具有已知体内效应的致畸剂和配对的非致畸剂。通过细胞蛋白含量、细胞活力(resazurin 还原和中性红摄取)和细胞收缩性的变化来测量差异效应。结果表明,致畸剂[L-蛋氨酸、5-氟尿嘧啶和磺胺异恶唑]可以与非致畸剂[DL-蛋氨酸、6-甲基尿嘧啶和磺胺分别区分开来。二氯醋酸和二氯丙醇影响胚胎干细胞,但不影响微团;二氯丙醇的影响大于二氯醋酸。这种方法揭示了对收缩性的差异影响,而不影响活性/活力,同时总细胞蛋白保持不变。我们建议应检查该系统的预验证。