Evaluation of human in vitro skin organ culture in a quasi-static planar homogeneous flow field

• Main Authors: YAN Hongtao, TANG Hui, QIU Weiming
• Format: Article
• Language: zho
• Published: Editorial Office of Journal of Third Military Medical University 2019-12-01
• Series: Di-san junyi daxue xuebao
• Subjects: skin culture; dynamic perfusion platform; in vitro culture; histomorphology
• Online Access: http://aammt.tmmu.edu.cn/Upload/rhtml/201907031.htm

Objective To observe and evaluate the effect of different perfusion frequencies on the survival of human skin specimens in a quasi-static planar homogeneous flow field. Methods The foreskin skin grafts in size of 4 mm were placed a quasi-static planar homogeneous flow field, and randomly divided into experimental group (perfused with high frequency) and the control group (perfused with low frequency). After 12 days' culture, PAS staining and VG staining were used to observe the changes of basement membrane, cell morphology and dermal collagen fibers. Moreover, the number of total cells and the apoptotic cells in the 2 groups were separately determined by HE staining and TUNEL assay. The dissolved oxygen concentration in culture medium was measured by a dissolved oxygen meter. Results In the low frequency perfusion group, gross observation showed characteristic features of necrosis; PAS and VG staining results indicated that the epidermis was severely separated, basement membrane structure disappeared and collagen fibers accumulated. Significant differences were seen in total cell number (249.35±13.00 vs 37.50±9.18, P < 0.001) and apoptotic rate between the experimental group and the control group. The dissolved oxygen level was higher in the former group than the latter group. Conclusion In the quasi-static planar homogeneous flow field, high frequency perfusion is suitable for the culture of free skin grafts in vitro.