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开发一种改进的气管外植体生物测定法以检测囊性纤维化血清中的纤毛运动障碍因子。

Development of an improved tracheal explant bioassay for the detection of the ciliary dyskinesia factor in cystic fibrosis serum.

作者信息

Gabridge M G, Bright M J, Agee C C, Nickerson J M, Henderson N S

出版信息

Pediatr Res. 1979 Jan;13(1):31-5. doi: 10.1203/00006450-197901000-00007.

Abstract

A tracheal ring explant system, when used with 25% cystic fibrosis (CF) serum, displayed obvious ciliostasis. Hamster, rabbit, and guinea pig explants all had measurable decreases in ciliary activity after 24 hr of incubation in the serum. The differential response to CF serum (relative to normal serum) was greatly increased by using explants which were maintained 24-72 hr in minimal essential medium (MEM) with 10% horse serum and which were selected on the basis of optimal ciliary activity and vigor. With such a bioassay system of guinea pig tracheal explants, incubation with 25% normal serum would produce essentially no change in relative ciliary activity (score of 242 of a possible 300), whereas CF serum resulted in an 86% decrease (score of 33). Scanning electron microscopic observation indicated that the explants displaying the CF-ciliostatic effect had significant accumulations of mucous over the ciliated epithelial surface. A biochemical viability assay (dehydrogenase activity) showed no cytonecrosis when CF serum-treated tissues were compared to standard explants (10% horse serum in MEM) or control explants (25% normal human serum).

摘要

气管环外植体系统与25%的囊性纤维化(CF)血清一起使用时,表现出明显的纤毛运动停滞。仓鼠、兔子和豚鼠的外植体在血清中孵育24小时后,纤毛活性均有可测量的下降。通过使用在含有10%马血清的最低必需培养基(MEM)中维持24 - 72小时,并根据最佳纤毛活性和活力选择的外植体,对CF血清(相对于正常血清)的差异反应大大增强。在这样的豚鼠气管外植体生物测定系统中,用25%的正常血清孵育,相对纤毛活性基本不会改变(可能的300分中得242分),而CF血清则导致下降86%(得33分)。扫描电子显微镜观察表明,表现出CF纤毛运动抑制作用的外植体在纤毛上皮表面有大量黏液积聚。生化活力测定(脱氢酶活性)显示,与标准外植体(MEM中含10%马血清)或对照外植体(25%正常人血清)相比,CF血清处理的组织没有细胞坏死。

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