人乳腺癌相关成纤维原代细胞培养的改良方法-肿瘤防治研究.PDF

2018 45 10  Cancer Res Prev Treat,2018,Vol.45,No.10 717 肿瘤防治研究 年第 卷第 期 · · doi:10.3971/j.issn.1000-8578.2018.18.0428 · · 基础研究 人乳腺癌相关成纤维原代细胞培养的改良方法 1,2 1,2 3 李智勇 ，白雪 ，蔡慧云 An Modified Primary Culture Method of Breast Cancer-associated Fibroblasts LI Zhiyong1,2, BAI Xue1,2, CAI Huiyun3 1. Clinical College of The PLA Army General Hospital, Anhui Medical University, Beijing 100700, China; 2. Department of Breast Surgery, The PLA Army General Hospital, Beijing 100700, China; 3. Center of General Surgery of PLA, The PLA Army General Hospital, Beijing 100700, China Corresponding Author: BAI Xue, E-mail: drbaixue@126.com Abstract: Objective To establish a rapid and high efficient method of primary culture of BCAFs by improving primary culture methods of common breast cancer-associated fibroblasts (BCAFs). Methods We took cancerous tissue samples from the patients diagnosed as invasive breast cancer. All samples were randomly divided into two groups: the modified group and the conventional group with same size and quality. The modified group was digested with a modified enzyme digestion method (type collagenase+hyaluro Ⅱ nidase digestion for 10 min). The conventional group was digested with conventional enzymatic digestion method (type collagenase digestion for 8h). The total number of cells obtained, the activity of adherent Ⅰ cells after 48h culture and the purity of BCAFs between two groups were compared by cell counting, MTT and immunofluorescence. The characteristics of primary BCAFs were observed under inverted microscope. Results The amount of cells,