Retrovirus-mediated transduction of porcine keratinocytes in organ culture.
Retrovirus-mediated transduction of porcine keratinocytes in organ culture.
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器官培养中逆转录病毒介导的猪角质形成细胞转导。
DOI：
10.1046/j.1523-1747.1998.00298.x
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发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Taichman,LB
中科院分区：
文献类型：
--
作者：
Ghazizadeh,S;Harrington,R;Garfield,J;Taichman,LB
将新的遗传信息直接转移到表皮的角质形成细胞中可能证明对治疗某些遗传性皮肤病是有效的;然而，目前的体内基因转移到皮肤的方法不会导致转基因的持续存在。本研究的目的是探索利用逆转录病毒介导的转导进行直接基因转移。逆转录病毒载体将其基因组的DNA拷贝整合到宿主染色体中，因此具有实现永久基因治疗的潜力。为了促进逆转录病毒载体体内转导方法的发展，构建了猪皮肤器官培养模型，其中裸露的表面用来自毛囊和表皮残余物的复制角质形成细胞重新填充。MFGlacZ（107个蓝色形成单位/ml）和用水泡性口炎病毒（VSV）G蛋白假型化的LZRN（109个菌落形成单位/ml），每种编码β-半乳糖苷酶报告基因，后者编码新霉素磷酸转移酶选择基因。与MFGlacZ相比，LZRN更频繁地观察到β-半乳糖苷酶表达细胞;然而，在两种情况下，转导效率仍然较低。在相同滴度下，VSV-G假型逆转录病毒载体显示出比具有嗜中性包膜的类似载体更有效地转染猪角质形成细胞。用G418原位筛选LZRN转导的细胞可增加器官培养中β-gal+细胞的数量。为了实现表皮细胞的体内转导，这些研究指出了高滴度逆转录病毒载体、VSV-G蛋白假型化和原位选择的重要性。
Direct transfer of new genetic information to keratinocytes in epidermis may prove effective in treating certain genodermatoses; however, current methods forin vivogene transfer to skin do not lead to persistence of the transgene. The goal of this study was to explore direct gene transfer using retrovirus-mediated transduction. Retroviral vectors integrate a DNA copy of their genome into the host chromosome and therefore have the potential to effect a permanent gene therapy. To facilitate development of methods forin vivotransduction with retroviral vectors, a porcine skin organ culture model was constructed in which the denuded surface was repopulated with replicating keratinocytes from hair follicles and epidermal remnants.In situtransduction was carried out by topical application of two retrovirus vectors, MFGlacZ (107blue forming units per ml) and LZRN pseudotyped with the G protein of vesicular stomatitis virus (VSV) (109colony forming units per ml), each encoding the β-galactosidase reporter gene and the latter encoding the neomycin phosphotransferase selectable gene. β-galactosidase expressing cells were observed more frequently with LZRN than with MFGlacZ; however, transduction efficiency remained low in both instances. At equivalent titers, the VSV-G pseudotyped retroviral vector was shown to transduce porcine keratinocytes more efficiently than a similar vector with the amphotropic envelope. The number of β-gal+ cells in organ culture could be increased by selection of LZRN-transduced cellsin situwith G418. To achieve transduction of epidermisin vivo, these studies point out the importance of high titer retroviral vectors, pseudotyping with VSV-G protein, andin situselection.
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DOI：
10.1111/1523-1747.ep12874554
发表时间：
1990-02
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
S. Regauer;C. Compton
通讯作者：
S. Regauer;C. Compton
影响因子：
56.9
作者：
E. Nabel;G. Plautz;G. Nabel
通讯作者：
E. Nabel;G. Plautz;G. Nabel
慢性伤口皮肤等效模型中逆转录病毒介导的基因转移。
影响因子：
4.6
作者：
Badiavas,E;Mehta,PP;Falanga,V
通讯作者：
Falanga,V
逆转录病毒介导的培养表皮角质形成细胞的转导。
DOI：
10.1111/1523-1747.ep12489019
发表时间：
1991
期刊：
The Journal of investigative dermatology
影响因子：
--
作者：
Garlick,JA;Katz,AB;Fenjves,ES;Taichman,LB
通讯作者：
Taichman,LB
移植逆转录病毒生产细胞，将体内基因转移到小鼠骨骼肌中。
影响因子：
4.2
作者：
A. Fassati;D. Wells;F. Walsh;G. Dickson
通讯作者：
G. Dickson