玻璃化保存工程化组织产品的实验与技术

Vitreous cryopreservation of engineered tissue bioproducts

目的:总结近年来玻璃化保存技术的新进展和多种工程化组织玻璃化保存的实验研究。探讨影响工程化组织低温保存的主要因素、评价指标等,以期指导玻璃化保存工程化组织的实验研究。资料来源:应用计算机检索Medline,EBSCO,ScienceDirect Onsite以及google scholar引擎中1980-02/2006-05关于细胞和组织低温保存的文献,检索词“tissue engineering,engineered tissue,cryopreservation,vitrification,ice-free cryopreservation,vitreous cryopreservation”,检索词被分别组合进行检索,限定文献语言种类为English。同时计算机检索万方数据库1980-02/2006-05关于细胞和组织低温保存的文献,检索词“组织工程,工程化组织,冻存,玻璃化保存”,限定文献语言种类为中文。资料选择:对资料进行初审,选出与研究关系密切的文献。纳入标准:研究对象为动物或人,包括基础与临床研究内容。重点选取与玻璃化保存技术基本原理相关以及具体的工程化构建物玻璃化保存相关的文献。排除标准:关于配子,胚胎和植物,食品等的玻璃化保存文章。资料提炼:共检索到122篇关于细胞和组织低温保存的文献,其中与本实验关系密切的文献17篇,间接相关的文献42篇,最终纳入31篇符合标准的文献。资料综合:组织工程产业化要求长期而稳定的保存包含活细胞的工程化组织产品,以及产品中细胞与材料的黏附。相对于传统的低温冻存技术而言,玻璃化保存因在保存过程中无细胞内外冰晶形成,可以改善复温后的细胞活力,并能保护细胞和支架材料之间的黏附。本文从近年来玻璃化保存技术的新进展和多种工程化组织玻璃化保存的实验研究进行综述,分析了玻璃化保存液要求高浓度,对细胞有非特异毒性损伤,对悬浮单细胞的玻璃化保存能得到较高的细胞存活,而与支架材料结合的种子细胞存活除受到冻存试剂和流程的影响外还受到自身结构的影响。结论:玻璃化保存是较好的生物系统保存方法,可以为工程化组织产品提供可靠的保存,但急需发现新的低毒高效的玻璃化试剂以及优化保存流程。

OBJECTIVE： To summarize the basic technological advances of vitreous cryopreservation and the experimental research of the vitreous cryopreservation of various engineered tissues, and explore the main factors, evaluated indexes influencing the vitreous cryopreservation, so as to guide the expedment of vitreous cryopreservation. DATA SOURCE, S： A computer-based online search of Medline, EBSCO, ScienceDirect Onsite and google scholar based search was undertaken to identify the articles on cells and tissues cryopreservation published in English from February 1980 to May 2006 with the key words of ＂tissue engineering, engineered tissue, cryopreservation, vitrification, ice-free cryopreservation, vitreous cryopreservation＂. Meanwhile, we searched Wanfang database for the related articles published in Chinese from February 1980 to May 2006 with the key words of ＂tissue engineedng, engineered tissue, cryopreservation, vitreous cryopreservation＂ in Chinese. STUDY SELECTION： The articles were pdmarily screened to select those closely related with the research. Inclusive criteda： the subjects should be animals or human; the study should be basic and/or clinical research on vitrification technology and expedments of some engineered tissue preservation. Exclusive cdteda： articles about the vitreous cryopreservation of gamete, embryo, plants and food. DATA EXTRACTION： Totally 122 articles about cells and tissue crydpreservation were found in these databases, among which 17 were closely associated with the study and 42 were indirectly related. Finally, 31 articles were selected for analysis. DATA SYNTHESIS： The commercialization of tissue engineedng needs long-term and stable preservation of engineered tissue bioproducts containing living cells and their adhesion to scaffolds. In contrast with the conventional freezing preservation method, vitreous cryopreservation improves the post-rewarming viability of cells and reduces intracellular and intercellular ice formation, therefore could protect the cell-scaffold adhesion. In this paper, we review the recent advances in vitreous cryopreservation and the vitreous cryopreservation of vadous engineered tissue, suggesting that vitreous .cryopreservation needs high concentration cryoprotectant, which is nonspecific toxic for cells; vitreous cryopreservation of cells suspension could preserve good cell survival; the viability of cells adhedng to the scaffold is determined by vitrification solution, protocols and tissue structure charactedstics. CONCLUSION： Vitreous cryopreservation is a good preserving method for the living system, and can offer reliable preservation for engineered tissues. But it is necessary to find new low-toxicity and high performance vitrification agents and preserving protocols.