摘要
目的尝试应用淋巴细胞分离液收集纯度高、活性强的上皮细胞进行体外培养,探讨是否可以加快上皮细胞培养的速度。方法根据上皮细胞和成纤维细胞比重上的差异,将经过胰酶处理后的小白鼠皮肤,用镊子把上皮和真皮层撕开,收集两层之间的基底细胞,并把这些细胞悬液加入淋巴细胞分层液(LP)中,收集中间层的细胞作为实验组进行体外培养,以传统方法分离的细胞培养作为对照组。两组同时进行对细胞生长速度的三种测试:上皮细胞生长曲线测定、Lowry蛋白测定和H^3-胸腺嘧啶核甙测定。结果经统计学分析证明,实验组的细胞生长速度与对照组比较有明显加快的表现。结论淋巴细胞分层液能获得活力强、生长快的细胞,可缩短培养成上皮细胞皮片的时间,有一定的临床意义。
Objective This study was to improve the rate of skin keratinocyte culture by applica- tion of a lymphocyte isolation fluid. Methods A lymphocyte isolation fluid 'Lymphoprep' contains 5.6% (W/V) of Ficoll, with a density of 1. 077g/ml. According to the difference between the densities of ker- atinocytes (1. 065-1. 085g/ml) and fibroblasts (1. 025-1. 050g/ml),the two ceil types can be separated by suspension in the solution. We performed a controlled study in new born mice: the epidermis and der- mis of skin were separated by forceps after trypsinization. The cell suspension collected was laid over Lymphoprep after centrifugation at 23℃, 8000 g for 10 min. The keratinocytes were collected at the middle-interphase, and these cells were pure and viable keratinocyte ceils. Results In subsequent ker- atinocyte culture we found that the cell growth was faster than cells obtained from conventional tech- nique. It greatly shortened the preparation time of keratinocyte cell sheet. Conclusion The method will be valuable clinically for treatment of burn patients.
