Retroviral transduction of murine epidermal stem cells demonstrates clonal units of epidermal structure
Retroviral transduction of murine epidermal stem cells demonstrates clonal units of epidermal structure
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DOI：
10.1111/1523-1747.ep12336255
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发表时间：
1997-09-01
影响因子：
6.5
通讯作者：
Mackenzie, IC
中科院分区：
文献类型：
--
作者：
Mackenzie, IC
已经提出表皮干细胞的数量和位置与上表皮层中的柱状结构单元有关，并且每个单元的细胞来源于单个干细胞。通过使用逆转录病毒转导提供组织化学上可证实的外源基因产物的遗传表达，促进了发育组织中细胞谱系的研究。为了提供关于表皮单位的克隆性质的直接证据，用携带β-半乳糖苷酶基因的复制缺陷型逆转录病毒载体转导小鼠表皮角质形成细胞。在体内表皮下注射病毒导致不频繁的转导，表皮内仅短暂存在β-galstaining角质形成细胞。角质形成细胞在体外转导和移植回体内部位允许在整个12周期间的改造表皮内的细胞簇中的转导基因的证明。表皮重新发育成有序的柱状结构，转导细胞局限于单个柱状单位。这种克隆外观与从单个干细胞衍生每个表皮单位相容，但与细胞增殖的随机模式不相容。与口腔粘膜结缔组织重组的转导的表皮片也重建了正常的柱状结构，β-gal染色限制于单个柱状单位。这些数据表明，与结构单位相关的表皮干细胞模式的建立是上皮的固有特性，并且不依赖于区域特异性结缔组织的影响。
It has been suggested that the number and position of epidermal stem cells are related to the units of columnar structure in the upper epidermal strata and that the cells of each unit are derived from a single stem cell. Studies of cell lineage in developing tissues have been facilitated by the use of retroviral transduction to provide inherited expression of a histochemically demonstrable foreign gene product. To provide direct evidence about the clonal nature of epidermal units, murine epidermal keratinocytes were transduced with a replication-deficient retroviral vector carrying the beta-galactosidase gene. Subepidermal injection of virus in vivo led to infrequent transduction with only transient presence of beta-galstaining keratinocytes within the epidermis. Transduction of keratinocytes in vitro and transplantation back to in vivo sites permitted demonstration of the transduced gene in clusters of cells within the reformed epidermis throughout a 12-wk period. The epidermis redeveloped an ordered columnar structure with restriction of transduced cells to individual columnar units. This clonal appearance is compatible with derivation of each epidermal unit from a single stem cell but is not compatible with a random pattern of cell proliferation. Transduced epidermal sheets that were recombined with oral mucosal connective tissue also redeveloped normal columnar structure with restriction of beta-gal staining to individual columnar units, These data suggest that the establishment of an epidermal stem cell pattern related to units of structure is an intrinsic property of the epithelium and is not dependent on regionally-specific connective tissue influences.