Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy
Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy
复制标题
DOI：
10.1002/jgm.740
复制
发表时间：
2005-07-01
影响因子：
3.5
通讯作者：
Mitrani, E
中科院分区：
文献类型：
--
作者：
Hasson, E;Slovatizky, Y;Mitrani, E
背景器官碎片可以在体外培养数周，如果它们被制备成显微镜厚度，并且如果基本器官结构被保留。这种器官片段，我们称之为微器官（MO），在培养中表达内源性组织特异性基因产物。我们已经利用这种方法来工程MOs离体通过基因transfer.Methods从脾，肺，结肠和皮肤制备的MOs感染使用：单纯疱疹病毒1型，腺病毒，牛痘病毒和小鼠白血病病毒（MuLV），携带报告基因β-半乳糖苷酶。优化后，在体外和植入同基因宿主后，使用MuLV构建体也获得了与用腺构建体获得的表达水平相当的高表达水平（> 15%阳性细胞）。植入后，工程组织被发现保持本地化，成为血管化，并表达转导的基因为几个months.Conclusions该系统可用于研究病毒和组织之间的相互作用，在体外和体内。此外，该方法为离体基因治疗提供了一个新的平台。这种工程结构可用作自体生物泵，用于在体内连续分泌具有临床重要性的基因产物。版权所有（c）2005年约翰威利父子有限公司。
Background Organ fragments can be cultured for weeks in vitro if they are prepared of microscopic thickness and if the basic organ structure is preserved. Such organ fragments, which we termed micro-organs (MOs), express in culture endogenous tissue-specific gene products. We have exploited this methodology to engineer MOs ex vivo by gene transfer.Methods MOs prepared from spleen, lung, colon and skin were infected using: herpes simplex type-1, adeno virus, vaccinia virus and murine leukemia virus (MuLV), carrying the reporter gene beta-galactosidase.Results All four viral vectors infected MOs in culture, with adeno infection giving significantly higher values. After optimization, high levels of expression (> 15% positive cells), comparable to those obtained with the adeno construct, were also obtained using the MuLV construct both in vitro and after implantation into syngeneic hosts. After implantation, the engineered tissue was found to remain localized, become vascularized, and to express the transduced gene for several months.Conclusions The system can be used to study interactions between viruses and tissues both ex vivo and in vivo. Furthermore, the approach proposes a novel platform for ex vivo gene therapy. Such engineered structures could be used as autologous biological pumps for continuous secretion in vivo of gene products of clinical importance. Copyright (c) 2005 John Wiley & Sons, Ltd.