Carlson Mark A, Prall Amy K, Gums Jeremiah J
Department of Surgery, VA Nebraska-Western Iowa Health Care System, Surgery 112, VA Medical Center, 4101 Woolworth Avenue, Omaha, NE 68105, USA.
Mol Cell Biochem. 2007 Dec;306(1-2):123-32. doi: 10.1007/s11010-007-9561-z. Epub 2007 Jul 27.
The technique of RNA interference (RNAi) was trialed in primary human foreskin fibroblasts, both in monolayer culture and in the fibroblast-populated collagen matrix. Knockdown of lamin A/C, p53, and FAK was possible with low-confluency (<50%) monolayer fibroblasts, a transfection vehicle concentration of 1%, and an siRNA concentration of 25-50 nM. Knockdown also was possible in the collagen matrix using similar reagent concentrations and a cellular density of one million fibroblasts per ml of matrix. Optimization of transfection conditions appeared to be important to increase knockdown efficiency. Consistent with prediction, knockdown of FAK induced apoptosis in the fibroblast-populated collagen matrix.
RNA干扰（RNAi）技术在原代人包皮成纤维细胞中进行了试验，包括单层培养和在成纤维细胞填充的胶原基质中。对于低汇合度（<50%）的单层成纤维细胞、1%的转染载体浓度和25 - 50 nM的小干扰RNA（siRNA）浓度，可实现对核纤层蛋白A/C、p53和黏着斑激酶（FAK）的敲低。使用相似的试剂浓度和每毫升基质中一百万个成纤维细胞的细胞密度，在胶原基质中也可实现敲低。优化转染条件对于提高敲低效率似乎很重要。与预测一致，在成纤维细胞填充的胶原基质中，FAK的敲低诱导了细胞凋亡。
