Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips.
Fabrication of Size-Controllable and Arrangement-Orderly HepG2 Spheroids for Drug Screening via Decellularized Liver Matrix-Derived Micropattern Array Chips.
复制标题
通过脱细胞肝基质衍生的微图案阵列芯片制备用于药物筛选的尺寸可控和排列有序的HepG2球体。
DOI：
10.1021/acsomega.1c06302
复制
发表时间：
2022-01-18
期刊：
影响因子：
4.1
通讯作者：
Bao J
中科院分区：
文献类型：
--
作者：
Zhu X;Wu Q;He Y;Gao M;Li Y;Peng W;Li S;Liu Y;Zhang R;Bao J
通过微阵列的三维（3D）培养，以产生细胞球体似乎是一个很有前途的体外仿生系统的肝组织工程应用，如药物筛选。最近，器官来源的脱细胞细胞外基质出现可以说是最仿生的生物墨水。本研究利用脱细胞肝基质（DLM）微阵列芯片制备尺寸可控、排列有序的HepG2球状体，用于药物筛选。猪DLM通过去除细胞成分获得，然后研磨成粉末，然后粉碎。在增强细胞粘附、增殖和功能的生物学性能方面，将DLM作为涂层基质与I型胶原（Col I）和Matrigel进行比较。随后，我们使用聚二甲基硅氧烷（PDMS）吸附DLM作为生物墨水来制作微图案阵列芯片。通过评估HepG2 3D细胞聚集体的形态、活力和功能来确定微图案的最佳形状和尺寸。此外，在该新型平台上进行了药物敏感性试验（紫杉醇、盐酸多柔比星和双硫仑）。与Col I和Matrigel相比，DLM提供了为HepG2细胞提供合适支持的组织特异性微环境。直径为100 μm的圆形微图案是快速制备大规模、尺寸可控、排列有序、具有三维球体形状和高细胞活力的HepG2细胞聚集体的最佳工艺参数。药物筛选试验表明，药物的效果可以直接证明芯片上通过共聚焦显微镜测量球体的活力。我们提供了一个新的平台，用于大规模产生的HepG2球状体具有均匀的大小和排列，从而带来方便，减少错误，并增加重现性的快速药物发现荧光定量分析。这种方法可能适用于推进个性化医疗和药物发现。
Three-dimensional (3D) culture via micropattern arrays to generate cellular spheroids seems a promising in vitro biomimetic system for liver tissue engineering applications, such as drug screening. Recently, organ-derived decellularized extracellular matrix emerges as arguably the most biomimetic bioink. Herein, decellularized liver matrix (DLM)-derived micropattern array chips were developed to fabricate size-controllable and arrangement-orderly HepG2 spheroids for drug screening. The porcine DLM was obtained by the removal of cellular components and then ground into powder, followed by enzymolysis. DLM as a coating substrate was compared with collagen type I (Col I) and Matrigel in terms of biological performance for enhancing cell adhesion, proliferation, and functions. Subsequently, we used poly(dimethylsiloxane) (PDMS) to adsorb DLM as the bioink to fabricate micropattern array chips. The optimal shape and size of micropattern were determined by evaluating the morphology, viability, and functions of HepG2 3D cellular aggregates. In addition, drug-susceptibility testing (paclitaxel, doxorubicin HCl, and disulfiram) was performed on this novel platform. The DLM provided the tissue-specific microenvironment that provided suitable supports for HepG2 cells, compared to Col I and Matrigel. A circular micropattern with a diameter of 100 μm was the optimal processing parameter to rapidly fabricate large-scale, size-controllable, and arrangement-orderly HepG2 cellular aggregates with 3D spheroid’s shape and high cell viability. Drug screening testing showed that the effect of a drug could be directly demonstrated on-chip by confocal microscopy measuring the viability of spheroids. We provide a novel platform for the large-scale generation of HepG2 spheroids with uniform size and arrangement, thus bringing convenience, reducing error, and increasing reproducibility for a rapid drug discovery by fluorescence quantitative analysis. This methodology may be possible to apply in advancing personalized medicine and drug discovery.
登录
查看更多内容
影响因子：
9.7
作者：
Mattei, G.;Di Patria, V.;Ahluwalia, A.
通讯作者：
Ahluwalia, A.
影响因子：
5.8
作者：
Badileanu A;Mora-Navarro C;Gracioso Martins AM;Garcia ME;Sze D;Ozpinar EW;Gaffney L;Enders JR;Branski RC;Freytes DO
通讯作者：
Freytes DO
影响因子：
14
作者：
Hannachi, Imen Elloumi;Itoga, Kazuyoshi;Okano, Teruo
通讯作者：
Okano, Teruo
影响因子：
9
作者：
Byun, Hayeon;Lee, Yu Bin;Shin, Heungsoo
通讯作者：
Shin, Heungsoo
影响因子：
2.8
作者：
Miyamoto, Daisuke;Hara, Takuya;Nakazawa, Kohji
通讯作者：
Nakazawa, Kohji