Haynes J H, Krummel T M, Flood L C, Cohen I K, Diegelmann R F
Department of Surgery, Medical College of Virginia, Virginia Commonwealth University, Richmond 23298-0001.
J Invest Surg. 1990;3(4):349-55. doi: 10.3109/08941939009140361.
The fetal response to cutaneous injury has been investigated in a variety of models; most have studied the differences between fetal and adult healing mechanisms in vivo and in cell culture. Further disclosure of the cellular and biochemical events requires a model that can be manipulated to study single factors influencing fetal tissue repair without the complex interactions that occur in vivo, but in a system that more closely approximates normal skin than cell culture models. This paper presents a method for the organ culture of fetal skin and its advantages as a model to help elucidate fetal healing mechanisms. Skin sections (1 x 1 cm) excised from the backs of fetuses of New Zealand white rabbits on day 27 gestation (term = 31 days) were placed eccentrically in 65-mm culture dishes and fed daily with 2.5 mL of DMEM containing 10% fetal calf serum, antibiotics, and 10 mM ascorbic acid. A separate group, treated similarly, received 4-mm punch wounds to assess the in vitro response to wounding. The specimens were incubated at 37 degrees C in humidified room air on a rocker platform to provide alternate exposure of the skin to air and medium. Gross observation at 3 weeks showed cells extending into the central wound, indicating that viable cells were proliferating and/or migrating from the tissue. Skin was examined histologically and was viable over the 3-week period studied. Organ culture, by maintaining tissue in the natural extracellular matrix, allows cell-to-cell contact and communication to be maintained while allowing controlled environmental manipulation.(ABSTRACT TRUNCATED AT 250 WORDS)
人们已在多种模型中研究了胎儿对皮肤损伤的反应;大多数研究都聚焦于胎儿与成体在体内及细胞培养中的愈合机制差异。要进一步揭示细胞和生化事件,需要一个能够进行操控的模型,以便在不存在体内复杂相互作用的情况下,研究影响胎儿组织修复的单一因素,而且该系统要比细胞培养模型更接近正常皮肤。本文介绍了一种胎儿皮肤器官培养的方法及其作为模型的优势,有助于阐明胎儿的愈合机制。从妊娠27天(足月为31天)的新西兰白兔胎儿背部切取的皮肤切片(1×1厘米),偏心放置于65毫米培养皿中,每天用2.5毫升含10%胎牛血清、抗生素和10毫摩尔抗坏血酸的DMEM培养基喂养。另一组接受类似处理,用4毫米打孔器造成伤口,以评估体外对损伤的反应。标本在37℃、湿度饱和的室内空气中的摇床上孵育,使皮肤交替接触空气和培养基。3周后的大体观察显示,细胞向中央伤口延伸,表明有活力的细胞正在从组织中增殖和/或迁移。在所研究的3周内,对皮肤进行了组织学检查,结果表明皮肤具有活力。器官培养通过将组织维持在天然细胞外基质中,既能保持细胞间的接触和通讯,又能对环境进行可控操作。(摘要截短至250字)