Mapping the Cell-Surface N-Glycoproteome of Human Hepatocytes Reveals Markers for Selecting a Homogeneous Population of iPSC-Derived Hepatocytes.
Mapping the Cell-Surface N-Glycoproteome of Human Hepatocytes Reveals Markers for Selecting a Homogeneous Population of iPSC-Derived Hepatocytes.
复制标题
DOI：
10.1016/j.stemcr.2016.07.016
复制
发表时间：
2016-09-13
影响因子：
5.9
通讯作者：
Duncan, Stephen A.
中科院分区：
文献类型：
--
作者：
Mallanna, Sunil K.;Cayo, Max A.;Twaroski, Kirk;Gundry, Rebekah L.;Duncan, Stephen A.
当比较来自不同肝病患者的IPSC来源的肝细胞样细胞之间的肝脏表型时，细胞异质性可能会混淆解释。我们提出，通过荧光激活细胞分选(FACS)可以产生均一的细胞群。利用细胞表面捕获蛋白质组学，我们在肝细胞上共鉴定了300个糖蛋白。对IPSCs分化过程中表达谱的分析表明，SLC10A1、CLRN3和AADAC在肝细胞分化的最后阶段高度丰富。流式细胞仪对表达SLC10A1、CLRN3或AADAC的肝细胞样细胞进行纯化，发现有肝细胞特征的细胞被浓缩。此外，转录组分析显示，表达肝脏基因调控网络的细胞丰富，而表达多能干细胞网络的细胞枯竭。总之，我们报道了在原代肝细胞中表达的细胞表面N-连接糖蛋白的广泛目录，并鉴定了有助于纯化IPSC来源的肝细胞样细胞的同质群体的细胞表面蛋白。已鉴定的占据原代人肝细胞表面的N-连接糖蛋白SLC10A1、CLRN3和AADAC在∼上表达25%的IPSC来源的肝细胞FAC利用这些标记丰富了IPSC来源的肝细胞的肝脏特征SLC10A1分离的细胞提供了均一的肝细胞样细胞群大多数IPSC分化方案产生的肝细胞具有成熟的肝脏标志物的异质性表达，这使得复杂的特征和微妙的表型分析变得混乱。Duncan及其同事利用蛋白质组学、转录图谱和流式细胞术来识别肝细胞限制性细胞表面蛋白，以标记IPSC来源的肝细胞亚群，允许纯化具有丰富肝脏特性的IPSC来源的肝细胞，并将分化之间的变异性降至最低。
When comparing hepatic phenotypes between iPSC-derived hepatocyte-like cells from different liver disease patients, cell heterogeneity can confound interpretation. We proposed that homogeneous cell populations could be generated by fluorescence-activated cell sorting (FACS). Using cell-surface capture proteomics, we identified a total of 300 glycoproteins on hepatocytes. Analyses of the expression profiles during the differentiation of iPSCs revealed that SLC10A1, CLRN3, and AADAC were highly enriched during the final stages of hepatocyte differentiation. FACS purification of hepatocyte-like cells expressing SLC10A1, CLRN3, or AADAC demonstrated enrichment of cells with hepatocyte characteristics. Moreover, transcriptome analyses revealed that cells expressing the liver gene regulatory network were enriched while cells expressing a pluripotent stem cell network were depleted. In conclusion, we report an extensive catalog of cell-surface N-linked glycoproteins expressed in primary hepatocytes and identify cell-surface proteins that facilitate the purification of homogeneous populations of iPSC-derived hepatocyte-like cells. Identified N-linked glycoproteins occupying surface of primary human hepatocytes SLC10A1, CLRN3, and AADAC are expressed on ∼25% of iPSC-derived hepatocytes FACS with these markers enriches hepatic character in iPSC-derived hepatocytes SLC10A1 sorted cells provide homogeneous populations of hepatocyte-like cells Most iPSC differentiation protocols generate hepatocytes with heterogeneous expression of mature hepatic markers, which confounds analyses of complex traits and subtle phenotypes. Duncan and colleagues utilize proteomics, transcriptional profiling, and FACS to identify hepatocyte-restricted cell-surface proteins that mark a subpopulation of iPSC-derived hepatocytes, allow purification of iPSC-derived hepatocytes with enriched hepatic character, and minimize variability between differentiations.
登录
查看更多内容
影响因子：
5.2
作者：
Agarwal, Sadhana;Holton, Katherine L.;Lanza, Robert
通讯作者：
Lanza, Robert
影响因子：
8.7
作者：
LANIER, LL;ENGLEMAN, EG;HERZENBERG, LA
通讯作者：
HERZENBERG, LA
影响因子：
13.5
作者：
Cayo, Max A.;Cai, Jun;DeLaForest, Ann;Noto, Fallon K.;Nagaoka, Masato;Clark, Brian S.;Collery, Ross F.;Si-Tayeb, Karim;Duncan, Stephen A.
通讯作者：
Duncan, Stephen A.
影响因子：
64.8
作者：
Wilhelm, Mathias;Schlegl, Judith;Kuster, Bernhard
通讯作者：
Kuster, Bernhard
影响因子：
7.7
作者：
Yan H;Zhong G;Xu G;He W;Jing Z;Gao Z;Huang Y;Qi Y;Peng B;Wang H;Fu L;Song M;Chen P;Gao W;Ren B;Sun Y;Cai T;Feng X;Sui J;Li W
通讯作者：
Li W