GENETICALLY-MODIFIED KERATINOCYTES TRANSPLANTED TO WOUNDS RECONSTITUTE THE EPIDERMIS
GENETICALLY-MODIFIED KERATINOCYTES TRANSPLANTED TO WOUNDS RECONSTITUTE THE EPIDERMIS
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DOI：
10.1073/pnas.91.20.9307
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发表时间：
1994-09-27
影响因子：
11.1
通讯作者：
ERIKSSON, E
中科院分区：
文献类型：
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作者：
VOGT, PM;THOMPSON, S;ERIKSSON, E
将表达β-半乳糖苷酶基因或人生长激素（hGH）基因的正常和逆转录病毒转染的角质形成细胞悬液移植到约克郡猪的封闭室皮肤全层伤口中。移植后4周获得的连续皮肤活检的免疫染色显示移植的角质形成细胞的存活以及β-半乳糖苷酶的表达。转染的角质形成细胞首先见于伤口的新真皮部分，然后见于再生的基底表皮层，最后见于棘层的终末分化细胞。当角质形成细胞转染hGH基因移植到类似的伤口，hGH检测10天的伤口液。相比之下，hGH在体外检测47天。移植转染或正常角质形成细胞的伤口比未移植对照组更快地恢复上皮屏障功能（P < 0.05）。该研究证实了角质形成细胞悬浮液的成功移植，它们的表皮重建，以及它们的修复加速。此外，表达β-半乳糖苷酶或hGH的遗传工程化移植角质形成细胞的这种明显正常的掺入表明将表达治疗性蛋白质的其他基因引入伤口以有利地影响愈合的可能性。伤口液检测表达的肽提供了hGH基因成功转移的早期证明。
Normal and retrovirally transfected keratinocyte suspensions expressing either the beta-galactosidase gene or the human growth hormone (hGH) gene were transplanted into chamber-enclosed skin full-thickness wounds of Yorkshire pigs. Immunostaining of sequential skin biopsies obtained for 4 weeks after transplantation showed survival of the transplanted keratinocytes as well as expression of beta-galactosidase, Transfected keratinocytes were first seen in the neodermal portions of the wounds, then in the regenerating basal epidermal layer, and finally in the terminally differentiating cells of the stratum spinosum. When keratinocytes transfected with the hGH gene were transplanted into similar wounds, hGH was detected for 10 days in wound fluid. In contrast, hGH was detected in vitro for 47 days. Wounds transplanted with either transfected or normal keratinocytes restored the epithelial barrier function significantly faster than nontransplanted controls (P < 0.05). The study confirms the successful transplantation of keratinocyte suspensions, their reconstitution of the epidermis, and their acceleration of repair. Further, this apparently normal incorporation of genetically engineered transplanted keratinocytes expressing either beta-galactosidase or hGH suggests the possibility of introducing other genes expressing therapeutic proteins into wounds to favorably affect healing. Wound fluid detection of the expressed peptide provided early demonstration of successful transfer of the hGH gene.