Xu Hong, Shi Hong-can, Zang Wang-fu, Lu Dan
Department of Cardiothoracic Surgery, Shanghai Rui Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Cryobiology. 2009 Apr;58(2):225-31. doi: 10.1016/j.cryobiol.2008.12.009. Epub 2008 Dec 30.
Vitrification is a promising alternative to tissue preservation, in which the tissue is permeated with cryoprotective agents (CPAs) in order to circumvent the hazardous effects associated with ice formation. In this study, we evaluate the effect of vitreous cryopreservation of rabbit trachea, by comparing vitrification procedure with conventional computer-programmed slow freezing approaches. Harvested rabbit trachea were tailored and divided into groups and cryopreserved by vitrification and programmed freezing, respectively. The morphology and ultrastructure of the thawed tracheal fragments including HE dyes, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end-labeling (TUNEL) staining, transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were studied to assess the integrity of the tracheal fragments. Morphological studies demonstrated that both cryopreservation procedure retained the integrity of trachea, both epithelial cells, cilia and cartilage cells were in good shape. Compared with slow freezing methods, vitrification was less detrimental to cartilage cells and had a higher survival rate of chondrocytes and coverage of epithelium and cilia. Therefore, vitrification procedure can be a more satisfactory method to preserve trachea and the survival of chondrocytes in situ in cartilage tissue is adequate and respiratory epithelium is soundly present.
玻璃化是一种很有前景的组织保存方法，在该方法中，组织会被冷冻保护剂（CPA）渗透，以规避与冰晶形成相关的有害影响。在本研究中，我们通过将玻璃化程序与传统的计算机编程慢速冷冻方法进行比较，评估兔气管玻璃化冷冻保存的效果。收获的兔气管经过修整并分组，分别通过玻璃化和程序冷冻进行冷冻保存。对解冻后的气管片段进行形态学和超微结构研究，包括苏木精-伊红（HE）染色、末端脱氧核苷酸转移酶介导的dUTP-地高辛标记（TUNEL）染色、透射电子显微镜（TEM）和扫描电子显微镜（SEM），以评估气管片段的完整性。形态学研究表明，两种冷冻保存程序均保留了气管的完整性，上皮，上皮细胞、纤毛和软骨细胞形态良好。与慢速冷冻方法相比，玻璃化对软骨细胞的损害较小，软骨细胞存活率更高，上皮细胞和纤毛的覆盖率更高。因此，玻璃化程序可能是一种更令人满意的气管保存方法，软骨组织中软骨细胞的原位存活率足够，呼吸上皮完好存在。
