Serrato D, Nieto-Aguilar R, Garzón I, Roda O, Campos A, Alaminos M
Tissue Engineering Group, Department of Histology, University of Granada, Spain.
Histol Histopathol. 2009 Dec;24(12):1531-40. doi: 10.14670/HH-24.1531.
The purpose of this study was to compare the effects of five different cryopreservation protocols on the histology of bioengineered tissues. Although several artificial tissues have been developed to the date by tissue engineering, classical histological analysis methods and techniques must be optimized for these new tissues with special properties. The results of this study showed that the use of volatile solutions (formaldehyde, glutaraldehyde, glacial acetic acid and acetone) was not able to prevent the formation of large ice crystals that, in turn, can alter the structure of the artificial tissues. However, preincubation of the tissues in different concentrations of a carbon hydrate (glucose, maltose or trehalose) resulted in a better preservation of the tissue structure. We conclude that the best protocol that allows for an efficient analysis of the bioengineered tissues with very few artifacts is preincubation of the tissues in 0.300M or 0.400M trehalose for 30 or 120 min prior to OCT (optimal cutting temperature) embedding and cryosectioning. For all those reasons, we recommend the use of a cryoprotective agent before OCT embedding of human artificial tissues.
本研究的目的是比较五种不同冷冻保存方案对生物工程组织组织学的影响。尽管到目前为止组织工程已开发出多种人工组织，但经典的组织学分析方法和技术必须针对这些具有特殊性质的新组织进行优化。本研究结果表明，使用挥发性溶液（甲醛、戊二醛、冰醋酸和丙酮）无法防止大冰晶的形成，而大冰晶反过来会改变人工组织的结构。然而，将组织在不同浓度的碳水化合物（葡萄糖、麦芽糖或海藻糖）中进行预孵育，能更好地保存组织结构。我们得出结论，在进行OCT（最佳切割温度）包埋和冷冻切片之前，将组织在0.300M或0.400M海藻糖中预孵育30或120分钟，是能够对生物工程组织进行高效分析且伪像极少的最佳方案。基于所有这些原因，我们建议在对人类人工组织进行OCT包埋之前使用冷冻保护剂。
