White S J, Page S M, Margaritis P, Brownlee G G
Chemical Pathology Unit, Sir William Dunn School of Pathology, University of Oxford, UK.
Hum Gene Ther. 1998 May 20;9(8):1187-95. doi: 10.1089/hum.1998.9.8-1187.
A persistent obstacle that has hampered gene transfer experiments is the short-term nature of transgene expression in vivo. In this article we present evidence for sustained expression from primary human keratinocytes, using the retroviral vector MFG. Primary keratinocytes were transduced in culture with the MFG retroviral vector containing the coding region from factor IX cDNA. Transduced keratinocytes, which secreted on average 830 ng of factor IX/10(6) cells/24 hr in tissue culture, were used to form a bilayered skin equivalent and grafted onto nude mice under a silicone transplantation chamber. Between 0.1 and 2.75 ng of human factor IX per milliliter was found in mouse plasma for more than 1 year, suggesting that keratinocyte stem cells were both transduced and grafted. The results show, for the first time, that long-term expression is obtainable in retrovirally transduced keratinocytes after transplantation.
阻碍基因转移实验的一个长期存在的障碍是体内转基因表达的短期性。在本文中，我们提供了使用逆转录病毒载体MFG从原代人角质形成细胞持续表达的证据。在培养中用含有因子IX cDNA编码区的MFG逆转录病毒载体转导原代角质形成细胞。转导的角质形成细胞在组织培养中平均每10(6)个细胞/24小时分泌830 ng因子IX，用于形成双层皮肤等效物，并在硅胶移植室下移植到裸鼠身上。在小鼠血浆中发现每毫升0.1至2.75 ng的人因子IX超过1年，这表明角质形成干细胞既被转导又被移植。结果首次表明，移植后逆转录病毒转导的角质形成细胞可实现长期表达。
