Determination of DNA content as quality control in decellularized tissues:challenges and pitfalls  

作　　者：Charlot Philips Lisanne Terrie Ewout Muylle Lieven Thorrez 

机构地区：[1]Tissue Engineering Lab,Department of Development and Regeneration,KU Leuven Campus Kulak,8500 Kortrijk,Belgium

出　　处：《Regenerative Biomaterials》2025年第1期170-177,共8页再生生物材料(英文版)

基　　金：supported by the Research Foundation—Flanders[G0D3620N and 1133520N]。

摘　　要：Decellularized organs and tissues are emerging within the field ofregenerative medicine to meet the growing demand for organand tissue transplantation. Quality control of these acellular matrices prior to transplantation is of paramount importance to ensure the absence of an adverse reaction. In particular, thoroughevaluation of the DNA content is essential but also poses technical challenges. Therefore, in this study, we compared differentmethods for quantitative and qualitative evaluation of DNA content in native and decellularized skeletal muscle tissue to identifystrengths and weaknesses for each. Histological analysis revealedthat Feulgen staining is more sensitive and robust than the commonly used hematoxylin–eosin and 40,6-diamidino-2-phenylindole staining for detection of remaining nuclear material.Furthermore, gel electrophoresis allowed to identify the quality and length of remaining DNA fragments. The results of the quantitativeanalysis indicated that direct measurement of DNA content in tissue lysates is preferred over silica-based extraction methods, since thelatter resulted in the loss of small DNA fragments during extraction. Moreover, a weight loss correction factor should be implementedto take into account the impact of the decellularization on the extracellular matrix. With regard to the detection method, the resultsrevealed that a fluorescence-based approach is more accurate than the use of UV/VIS absorbance. Through combination of the proposedmethods, it should be possible to achieve a more standardized evaluation of novel acellular matrices in terms of DNA content and toenhance the predictability of clinical success.

关 键 词：DECELLULARIZATION skeletal muscle DNA quantification SPECTROPHOTOMETER 

分 类 号：R73[医药卫生—肿瘤]