Decellularized Liver Nanofibers Enhance and Stabilize the Long‐Term Functions of Primary Human Hepatocytes In Vitro
Decellularized Liver Nanofibers Enhance and Stabilize the Long‐Term Functions of Primary Human Hepatocytes In Vitro
复制标题
脱细胞肝脏纳米纤维增强并稳定体外原代人肝细胞的长期功能
DOI：
10.1002/adhm.202202302
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发表时间：
2023
影响因子：
10
通讯作者：
Khetani, Salman R.
中科院分区：
文献类型：
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作者：
Liu, Jennifer S.;Madruga, Liszt Y. C.;Yuan, Yang;Kipper, Matt J.;Khetani, Salman R.
由于不同物种肝脏功能的显著差异，体外人体肝脏模型被用于筛选化合物的新陈代谢和毒性，模拟疾病和基于细胞的治疗。然而，用于这类模型的细胞外基质(ECM)支架通常既不能模拟天然肝ECM的复杂成分，也不能模拟其纳米纤维的形态。因此，这里开发了一种新的方法，在没有合成聚合物共混物的情况下，将脱细胞猪肝细胞外基质和I型胶原蛋白电纺成纳米和微纤维(≈200-1000 nm)。原代人肝细胞(PHH)在纳米纤维上单独培养或与非实质细胞(3T3-J2胚胎成纤维细胞或原代人肝内皮细胞)共培养时，白蛋白分泌、尿素合成和细胞色素P450 1A2、2A6、2C9和3A4酶活性均高于常规吸附的ECM对照组。在成纤维细胞存在的近7周内，胶原/PLECM共混纳米纤维的PHH功能最高(与吸附的ECM相比，CYP3A4活性高出34倍)。综上所述，在体外数周内，首次显示细胞外基质的组成和形貌协同增强和稳定PHH的功能。纳米纤维平台可用于上述应用，并可用于阐明人体肝脏中细胞-ECM的相互作用。
Owing to significant differences across species in liver functions, in vitro human liver models are used for screening the metabolism and toxicity of compounds, modeling diseases, and cell‐based therapies. However, the extracellular matrix (ECM) scaffold used for such models often does not mimic either the complex composition or the nanofibrous topography of native liver ECM. Thus, here novel methods are developed to electrospin decellularized porcine liver ECM (PLECM) and collagen I into nano‐ and microfibers (≈200–1000 nm) without synthetic polymer blends. Primary human hepatocytes (PHHs) on nanofibers in monoculture or in coculture with nonparenchymal cells (3T3‐J2 embryonic fibroblasts or primary human liver endothelial cells) display higher albumin secretion, urea synthesis, and cytochrome‐P450 1A2, 2A6, 2C9, and 3A4 enzyme activities than on conventionally adsorbed ECM controls. PHH functions are highest on the collagen/PLECM blended nanofibers (up to 34‐fold higher CYP3A4 activity relative to adsorbed ECM) for nearly 7 weeks in the presence of the fibroblasts. In conclusion, it is shown for the first time that ECM composition and topography synergize to enhance and stabilize PHH functions for several weeks in vitro. The nanofiber platform can prove useful for the above applications and to elucidate cell‐ECM interactions in the human liver.