3D or not 3D: a guide to assess cell viability in 3D cell systems
3D or not 3D: a guide to assess cell viability in 3D cell systems
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DOI：
10.1039/d2sm00018k
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发表时间：
2022-02-15
期刊：
影响因子：
3.4
通讯作者：
Shpichka, Anastasia
中科院分区：
文献类型：
--
作者：
Bikmulina, Polina;Kosheleva, Nastasia;Shpichka, Anastasia
细胞活力是用于各种目的的主要综合参数，特别是当制造组织等同物（例如，大多数常规方法最初设计用于单层（2D）培养;然而，2D方法未能充分评估组织工程化构建体的存活力和药物作用，并且不能再现宿主-病原体相互作用和感染性。本研究旨在揭示特定的三维细胞系统的参数，如组分的浓度，凝胶厚度，细胞密度等对细胞活力和标准检测方法的适用性的影响。在这里，我们提出了一种方法，以实现充分的和可重复的结果上的细胞活力在3D胶原蛋白和纤维蛋白为基础的系统使用活/死，AlamarBlue和PicoGreen测定。我们的结果表明，应该根据不同的细胞特性（例如代谢活性、增殖能力、形态学等）使用至少三种方法的组合来进行3D系统的常规精确分析。
Cell viability is the primary integrative parameter used for various purposes, particularly when fabricating tissue equivalents (e.g., using bioprinting or scaffolding techniques), optimizing conditions to cultivate cells, testing chemicals, drugs, and biomaterials, etc. Most of the conventional methods were originally designed for a monolayer (2D) culture; however, 2D approaches fail to adequately assess a tissue-engineered construct's viability and drug effects and recapitulate the host-pathogen interactions and infectivity. This study aims at revealing the influence of particular 3D cell systems' parameters such as the components' concentration, gel thickness, cell density, etc. on the cell viability and applicability of standard assays. Here, we present an approach to achieving adequate and reproducible results on the cell viability in 3D collagen- and fibrin-based systems using the Live/Dead, AlamarBlue, and PicoGreen assays. Our results have demonstrated that a routine precise analysis of 3D systems should be performed using a combination of at least three methods based on different cell properties, e.g. the metabolic activity, proliferative capacity, morphology, etc.