Imaging-Based Screen Identifies Laminin 411 as a Physiologically Relevant Niche Factor with Importance for i-Hep Applications.
Imaging-Based Screen Identifies Laminin 411 as a Physiologically Relevant Niche Factor with Importance for i-Hep Applications.
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DOI：
10.1016/j.stemcr.2018.01.025
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发表时间：
2018-03-13
影响因子：
5.9
通讯作者：
Rashid ST
中科院分区：
文献类型：
--
作者：
Ong J;Serra MP;Segal J;Cujba AM;Ng SS;Butler R;Millar V;Hatch S;Zimri S;Koike H;Chan K;Bonham A;Walk M;Voss T;Heaton N;Mitry R;Dhawan A;Ebner D;Danovi D;Nakauchi H;Rashid ST
与原代细胞相比，诱导多能干细胞（i-Heps）衍生的肝细胞的使用受到其功能差异的限制。细胞外生态位因子可能在弥合这一差距中发挥关键作用。使用来自17名人类供体的新鲜分离的肝细胞的基于图像的表征（高含量分析; HCA），我们设计并验证了用于将细胞的肝脏特性与生理金标准进行比较的算法（肝细胞相似性指数; HLI）。然后将HLI应用于细胞外生态位因子的靶向筛选中，以鉴定驱动i-Heps更接近标准的底物。层粘连蛋白411（最高命中）在另外两个诱导多能干细胞（iPSC）系、原代组织和α1-抗胰蛋白酶缺乏症的体外模型中得到验证。累积起来，这些数据提供了一种参考方法来控制和筛选i-Hep分化，将层粘连蛋白411鉴定为关键的小生境蛋白，并强调了在开发iPSC应用时结合底物、可溶性因子和HCA的重要性。iPSC衍生的肝细胞（i-Hep）与原代细胞相比在功能上是有限的细胞外小生境内的因子可能在弥合这一差距中发挥作用层粘连蛋白411被证明是i-Hep的重要小生境因子高含量图像分析（HCA）可以帮助开发i-Hep应用Rashid和同事证明了高含量图像分析的实用性。本发明提供了用于鉴定生理学相关的细胞外小生境因子以使i-Heps更接近其主要组织对应物的通量成像平台。细胞外基质（ECM）蛋白筛选将层粘连蛋白411鉴定为促进基于i-Hep的体外疾病建模的重要生态位因子。
Use of hepatocytes derived from induced pluripotent stem cells (i-Heps) is limited by their functional differences in comparison with primary cells. Extracellular niche factors likely play a critical role in bridging this gap. Using image-based characterization (high content analysis; HCA) of freshly isolated hepatocytes from 17 human donors, we devised and validated an algorithm (Hepatocyte Likeness Index; HLI) for comparing the hepatic properties of cells against a physiological gold standard. The HLI was then applied in a targeted screen of extracellular niche factors to identify substrates driving i-Heps closer to the standard. Laminin 411, the top hit, was validated in two additional induced pluripotent stem cell (iPSC) lines, primary tissue, and an in vitro model of α1-antitrypsin deficiency. Cumulatively, these data provide a reference method to control and screen for i-Hep differentiation, identify Laminin 411 as a key niche protein, and underscore the importance of combining substrates, soluble factors, and HCA when developing iPSC applications. iPSC-derived hepatocytes (i-Heps) are functionally limited compared with primary cells Factors within the extracellular niche likely play a role in bridging this gap Laminin 411 was shown to be an important niche factor for i-Heps High content image analysis (HCA) can help development of i-Hep applications Rashid and colleagues demonstrate the utility of a high-throughput imaging platform for identification of physiologically relevant extracellular niche factors to advance i-Heps closer to their primary tissue counterparts. The extracellular matrix (ECM) protein screen identified Laminin 411 as an important niche factor facilitating i-Hep-based disease modeling in vitro.
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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：
48
作者：
Langmead, Ben;Salzberg, Steven L.
通讯作者：
Salzberg, Steven L.
影响因子：
3.7
作者：
Massey AJ
通讯作者：
Massey AJ
影响因子：
13.5
作者：
Rowe, Cliff;Gerrard, Dave T.;Jenkins, Roz;Berry, Andrew;Durkin, Kesta;Sundstrom, Lars;Goldring, Chris E.;Park, B. Kevin;Kitteringham, Neil R.;Hanley, Karen Piper;Hanley, Neil A.
通讯作者：
Hanley, Neil A.
影响因子：
4.8
作者：
Bouchecareilh, Marion;Hutt, Darren M.;Balch, William E.
通讯作者：
Balch, William E.