Chemiluminescent restriction fragment length polymorphism analysis (DNA fingerprinting) to identify keratinocytes from two donors in co-culture.
Chemiluminescent restriction fragment length polymorphism analysis (DNA fingerprinting) to identify keratinocytes from two donors in co-culture.
复制标题
化学发光限制性片段长度多态性分析（DNA 指纹分析）可鉴定来自共培养的两个供体的角质形成细胞。
DOI：
10.1111/j.1600-0625.1993.tb00195.x
复制
发表时间：
1993
影响因子：
3.6
通讯作者：
Cuono,CB
中科院分区：
文献类型：
--
作者：
Young,DM;Summers,WC;Cuono,CB
The transplantation of allogeneic cells such as pancreatic islet cells, bone marrow, and keratinocytes for skin replacement is in growing use clinically. Present techniques which are used to distinguish transplanted allogeneic cells from phenolypically identical host cells in chimeric tissue have serious limitations. This study assesses the feasibility of using Chemiluminescent restriction fragment length polymorphism (RFLP) analysis to distinguish between keratinocyte cell lines from 2 different donors grown in co‐culture. We evaluated the sensitivity of this technique in tissue cultured cells in order to define its usefulness in clinical situations. RFLP analysis readily detects heterologous DNA comprising only 10% of a 5 μg sample. This was a detection threshold of 500 ng of high molecular wefght DNA. With standard extraction methods, this represents the DNA obtained from 4000 cells. This new chemiluminescent technique is as sensitive as older techniques which employ radioactive probes, but avoids the hazards of handling radioactive material. Using this RFLP analysis, the presence of transplanted allogeneic cells can be readily delected in small biopsy specimens. This technique has wide potential application in allogeneic cellular transplantation investigations.
