[1]闫洪涛,唐辉,邱伟明,等.准静态平面均匀流场条件下游离皮片体外培养效果评价[J].第三军医大学学报,2019,41(23):2335-2340.
 YAN Hongtao,TANG Hui,QIU Weiming,et al.Evaluation of human in vitro skin organ culture in a quasi-static planar homogeneous flow field[J].J Third Mil Med Univ,2019,41(23):2335-2340.
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第23期
页码:
2335-2340
栏目:
基础医学
出版日期:
2019-12-15

文章信息/Info

Title:
Evaluation of human in vitro skin organ culture in a quasi-static planar homogeneous flow field
作者:
闫洪涛唐辉邱伟明宋林娟杨桂红伍津津
400042 重庆,陆军军医大学(第三军医大学)大坪医院皮肤科1;430071 武汉,中国人民解放军中部战区总医院皮肤整形美容科2
Author(s):
YAN Hongtao1 TANG Hui1 QIU Weiming2 SONG Linjuan1 YANG Guihong1 WU Jinjin1

1Department of Dermatology, Daping Hospital, Army Medical University (Third Military Medical University), Chongqing, 400042; 2Department of Plastic Surgery, General Hospital of Central Theater Command of PLA, Wuhan, Hubei Province, 430071, China

关键词:
皮肤组织器官自动灌注系统体外培养组织形态学
Keywords:
skin culture dynamic perfusion platform in vitro culture histomorphology
分类号:
R322.99; R329-33
文献标志码:
A
摘要:

目的观察并评价准静态平面均匀流场不同灌注频次对游离人皮肤标本的培养效果。方法将4 mm游离包皮皮片置于准静态平面均匀流场中,按随机数字表法分为2组:实验组采用高频次灌注,对照组采用低频次灌注;培养12 d后进行PAS染色、VG染色观察基底膜、细胞形态及真皮胶原纤维的变化情况;采用HE染色,TUNEL法观察2组皮片中细胞数目及细胞凋亡的变化情况;利用溶解氧测量仪模拟测量培养基中溶解氧的含量。结果肉眼观察见对照组皮片呈坏死特征性变化;PAS、VG染色结果显示对照组表真皮严重分离,基底膜结构消失,胶原纤维堆积;实验组染色细胞数为(249.35±13.00),显著高于对照组[(37.50±9.18),P<0.001];实验组细胞凋亡率为(9.91±3.89)%,显著低于对照组[(59.73±8.04)%,P<0.001];高频次灌注溶解氧水平高于低频次灌注。结论准静态平面均匀流场条件下,高频次灌注对人皮肤游离标本的体外培养作用更佳。

Abstract:

ObjectiveTo observe and evaluate the effect of different perfusion frequencies on the survival of human skin specimens in a quasi-static planar homogeneous flow field. MethodsThe 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. ResultsIn 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 [(9.91±3.89)% vs (59.73±8.04)%, P<0.001] between the experimental group and the control group. The dissolved oxygen level was higher in the former group than the latter group. ConclusionIn the quasi-static planar homogeneous flow field, high frequency perfusion is suitable for the culture of free skin grafts in vitro.

更新日期/Last Update: 2019-12-09