Shon Y H, Wolfinbarger L
Center for Biotechnology, Old Dominion University, Norfolk, Virginia 23529.
Cryobiology. 1994 Apr;31(2):121-32. doi: 10.1006/cryo.1994.1016.
Quantitative analysis of proteoglycans (PGs) revealed that the content of PG material from cryopreserved aorta, measured as uronate-positive material, was similar to that from fresh tissue (440 +/- 30 versus 430 +/- 7 micrograms/g wet tissue). Gel permeation column chromatography studies suggested that three PG fractions from cryopreserved tissue had molecular weights similar to PG fractions from fresh tissue; K(av) = 0.13, 0.47 (I), 0.20 (II), and 0.43 (III) from cryopreserved tissue and K(av) = 0.13, 0.50 (I), 0.23 (II), and 0.40 (III) from fresh tissue. Sequential extraction of tissue with guanidine-HCl (Gdn-HCl) followed by digestions with collagenase, elastase, and papain also demonstrated that there was no difference between fresh and cryopreserved tissues in the distribution of PGs in the extracts. Transmission electron microscopy analysis revealed less densely packed collagen fibers in cryopreserved tissues compared to fresh tissues. These studies indicate that there is no significant alteration in the content, molecular size, or distribution of PGs in properly cryopreserved tissue.
蛋白聚糖(PGs)的定量分析显示,以尿酸盐阳性物质衡量,冷冻保存主动脉中PG物质的含量与新鲜组织相似(440±30对430±7微克/克湿组织)。凝胶渗透柱色谱研究表明,冷冻保存组织中的三个PG组分的分子量与新鲜组织中的PG组分相似;冷冻保存组织的K(av) = 0.13、0.47(I)、0.20(II)和0.43(III),新鲜组织的K(av) = 0.13、0.50(I)、0.23(II)和0.40(III)。用盐酸胍(Gdn-HCl)对组织进行连续提取,然后用胶原酶、弹性蛋白酶和木瓜蛋白酶消化,也表明新鲜组织和冷冻保存组织提取物中PGs的分布没有差异。透射电子显微镜分析显示,与新鲜组织相比,冷冻保存组织中的胶原纤维排列不那么紧密。这些研究表明,在适当冷冻保存的组织中,PGs的含量、分子大小或分布没有显著改变。