The Microfluidic Environment Reveals a Hidden Role of Self-Organizing Extracellular Matrix in Hepatic Commitment and Organoid Formation of hiPSCs.
The Microfluidic Environment Reveals a Hidden Role of Self-Organizing Extracellular Matrix in Hepatic Commitment and Organoid Formation of hiPSCs.
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DOI：
10.1016/j.celrep.2020.108453
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发表时间：
2020-12-01
期刊：
影响因子：
8.8
通讯作者：
Elvassore N
中科院分区：
文献类型：
--
作者：
Michielin F;Giobbe GG;Luni C;Hu Q;Maroni I;Orford MR;Manfredi A;Di Filippo L;David AL;Cacchiarelli D;De Coppi P;Eaton S;Elvassore N
在人类肝脏发育过程中，肝脏身份的确定严格由外部信号控制，但细胞如何通过激活分泌级联反应这些外源信号仍不清楚，这是非常相关的，特别是在3D自组织系统中。在这里，我们研究了人类多能干细胞（hPSCs）在响应发育的外源信号时分泌的蛋白质如何影响从内胚层到肝脏谱系的进展，包括它们产生新生肝类器官的能力。在细胞培养耦合质谱（SILAC-MS）定量蛋白质组学分析中，通过微流体受限环境和氨基酸稳定同位素标记，我们发现了高丰度的细胞外基质（ECM）相关蛋白。来源于微流体的肝祖细胞或暴露于外源性ECM刺激下的肝祖细胞显示出明显更高的形成肝类器官的潜力，这些类器官可以快速扩增几代并进一步分化为功能性肝细胞。这些结果证明了对下游应用的肝类器官形成和分化效率的额外控制。基于SILAC的蛋白质组学分析显示高丰度分泌的ECM蛋白ECM沉积和重塑与细胞ECM受体过表达相关内源性或外源性ECM促进类器官的形成和分化Michielin等人通过将微流体与SILAC蛋白质组学分析相结合来研究人多能干细胞在肝脏分化过程中的分泌组。它们揭示了可溶性ECM蛋白积累和沉积的作用，并利用这些见解有效而稳健地从hipsc中获得肝类器官。
The specification of the hepatic identity during human liver development is strictly controlled by extrinsic signals, yet it is still not clear how cells respond to these exogenous signals by activating secretory cascades, which are extremely relevant, especially in 3D self-organizing systems. Here, we investigate how the proteins secreted by human pluripotent stem cells (hPSCs) in response to developmental exogenous signals affect the progression from endoderm to the hepatic lineage, including their competence to generate nascent hepatic organoids. By using microfluidic confined environment and stable isotope labeling with amino acids in cell culture-coupled mass spectrometry (SILAC-MS) quantitative proteomic analysis, we find high abundancy of extracellular matrix (ECM)-associated proteins. Hepatic progenitor cells either derived in microfluidics or exposed to exogenous ECM stimuli show a significantly higher potential of forming hepatic organoids that can be rapidly expanded for several passages and further differentiated into functional hepatocytes. These results prove an additional control over the efficiency of hepatic organoid formation and differentiation for downstream applications. Microfluidic confined environment enhances hepatic differentiation of hPSCs SILAC-based proteomic analysis reveals high abundance of secreted ECM proteins ECM deposition and remodeling correlate with cell-ECM receptor overexpression Either endogenous or exogenous ECM enhances organoid formation and differentiation Michielin et al. investigate the secretome of human pluripotent stem cells undergoing hepatic differentiation by coupling microfluidics with SILAC proteomic analysis. They reveal a role of soluble ECM protein accumulation and deposition and leverage these insights to efficiently and robustly derive hepatic organoids from hiPSCs.
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DOI：
10.1093/bioinformatics/btu638
发表时间：
2015-01-15
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Anders S;Pyl PT;Huber W
通讯作者：
Huber W
影响因子：
25.7
作者：
Godoy P;Schmidt-Heck W;Natarajan K;Lucendo-Villarin B;Szkolnicka D;Asplund A;Björquist P;Widera A;Stöber R;Campos G;Hammad S;Sachinidis A;Chaudhari U;Damm G;Weiss TS;Nüssler A;Synnergren J;Edlund K;Küppers-Munther B;Hay DC;Hengstler JG
通讯作者：
Hengstler JG
DOI：
10.1073/pnas.0914019107
发表时间：
2010-02-23
影响因子：
11.1
作者：
Gonzalez, Rodolfo;Jennings, Lori L.;Lesley, Scott A.
通讯作者：
Lesley, Scott A.
影响因子：
4.8
作者：
Barker, TH;Baneyx, G;Sage, EH
通讯作者：
Sage, EH
增强内源信号传导下用于分泌蛋白组分析的微流体
DOI：
10.1016/j.bbrc.2018.02.025
发表时间：
2018-03-04
影响因子：
3.1
作者：
Hu, Qianjiang;Luni, Camilla;Elvassore, Nicola
通讯作者：
Elvassore, Nicola