A resazurin-based,nondestructive assay for monitoring cell proliferation during a scaffold-based 3D culture process  

作　　者：Xianghui Gong Zhuqing Liang Yongxing Yang Haifeng Liu Jing Ji Yubo Fan 

机构地区：[1]Key Laboratory for Biomechanics and Mechanobiology of Ministry of Education,School of Biological Science and Medical Engineering,Beihang University,Beijing 100083,People’s Republic of China [2]Beijing Advanced Innovation Centre for Biomedical Engineering,Beihang University,Beijing 102402,People’s Republic of China [3]National Research Center for Rehabilitation Technical Aids,Beijing 100176,People’s Republic of China

出　　处：《Regenerative Biomaterials》2020年第3期271-281,共11页再生生物材料（英文版）

基　　金：supported by the National Natural Science Foundation of China(31771019,11172031,11827803,and 31470901);The National Key Research and Development Plan(2016YFC1101101);International Joint Research Center of Aerospace Biotechnology and 344 Medical Engineering from Ministry of Science and Technology of China,111 Project 345(B13003).

摘　　要：Development of viable cell estimation method without sacrificing proliferation and functions of cells cultured on regenerative biomaterials is essential for regenerative engineering.Cytotoxicity and depletion of resazurin are critical but often overlooked limitations that hindered applications of resazurin in viable cell estimation.The present work found that cytotoxicity and depletion of resazurin depended on cell concentration,resazurin concentration and resazurin incubation time.A simple strategy which only allowed cells to incubate with resazurin during each measurement was developed to eliminate negative effects of resazurin.This strategy was verified by monitoring proliferation of MC3T3-E1 preosteoblasts on poly(D,L-lactic acid)scaffold during a continuous 3D culture process for up to 21 days,comparing the accuracy with MTT assay which is a destructive assay with high sensitivity and accuracy and commonly used in regenerative engineering and comparing viability,proliferation and differentiation functions of MC3T3-E1,which were treated with/without this strategy for nondestructive evaluation.This method showed comparable linearity of standard curve and characteristics of growth curve to MTT assay.No major negative effects of this method on MC3T3-E1 viability and functions were found.Our work highlighted the importance of the concentration and incubation time of resazurin in designing application-specific nondestructive viability assay and would be helpful in improving the implanted medical devices as well as in regenerative engineering.

关 键 词：RESAZURIN nondestructive viable cell estimation CYTOTOXICITY resazurin depletion cell-based regenerative medicine 

分 类 号：R96[医药卫生—药理学]