Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion.
Identification of polymer surface adsorbed proteins implicated in pluripotent human embryonic stem cell expansion.
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DOI：
10.1039/c6bm00214e
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发表时间：
2016-08-16
影响因子：
6.6
通讯作者：
Alexander MR
中科院分区：
文献类型：
--
作者：
Hammad M;Rao W;Smith JG;Anderson DG;Langer R;Young LE;Barrett DA;Davies MC;Denning C;Alexander MR
利用高通量筛选发现热休克蛋白作为人类多能干细胞培养的候选物。需要改进的生物材料用于再生医学、生物传感和作为医疗器械。细胞对培养基中培养的聚合物的化学反应通常被认为是由吸附到表面的蛋白质主导的。在这里，我们使用质谱法，以确定从复杂的小鼠胚胎成纤维细胞（MEF）的条件培养基中吸附的蛋白质，发现支持多能性人胚胎干细胞（hESC）扩增等离子蚀刻组织培养聚苯乙烯表面。总共鉴定了71种蛋白质，其中14种与实现多能干细胞扩增的表面独特相关。我们已经开发了一种微阵列组合蛋白质点样方法来测试这14种蛋白质在新型聚合物（N-（4-羟苯基）甲基丙烯酰胺）上支持hESC细胞系（HUES-7）和人诱导多能干细胞系（ReBl-PAT）扩增的潜力。这些蛋白质被点样以形成一个初级阵列，产生几个蛋白质混合物“命中”，增强细胞附着到聚合物。生成第二阵列以测试一组精制蛋白质混合物的功能。我们发现，热休克蛋白90和热休克蛋白-1的组合鼓励多能干细胞的粘附性提高到与纤连蛋白预处理相当的水平。
The discovery of heat shock proteins as candidates for human pluripotent stem cell culture using high throughput screening. Improved biomaterials are required for application in regenerative medicine, biosensing, and as medical devices. The response of cells to the chemistry of polymers cultured in media is generally regarded as being dominated by proteins adsorbed to the surface. Here we use mass spectrometry to identify proteins adsorbed from a complex mouse embryonic fibroblast (MEF) conditioned medium found to support pluripotent human embryonic stem cell (hESC) expansion on a plasma etched tissue culture polystyrene surface. A total of 71 proteins were identified, of which 14 uniquely correlated with the surface on which pluripotent stem cell expansion was achieved. We have developed a microarray combinatorial protein spotting approach to test the potential of these 14 proteins to support expansion of a hESC cell line (HUES-7) and a human induced pluripotent stem cell line (ReBl-PAT) on a novel polymer (N-(4-Hydroxyphenyl) methacrylamide). These proteins were spotted to form a primary array yielding several protein mixture ‘hits’ that enhanced cell attachment to the polymer. A second array was generated to test the function of a refined set of protein mixtures. We found that a combination of heat shock protein 90 and heat shock protein-1 encourage elevated adherence of pluripotent stem cells at a level comparable to fibronectin pre-treatment.
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DOI：
10.1002/adma.201501351
发表时间：
2015-07-15
期刊：
Advanced materials (Deerfield Beach, Fla.)
影响因子：
--
作者：
Celiz AD;Smith JG;Patel AK;Hook AL;Rajamohan D;George VT;Flatt L;Patel MJ;Epa VC;Singh T;Langer R;Anderson DG;Allen ND;Hay DC;Winkler DA;Barrett DA;Davies MC;Young LE;Denning C;Alexander MR
通讯作者：
Alexander MR
影响因子：
64.5
作者：
Mitsui, K;Tokuzawa, Y;Yamanaka, S
通讯作者：
Yamanaka, S
DOI：
10.1016/j.bbagen.2013.03.019
发表时间：
2013-06-01
影响因子：
3
作者：
Sheokand, Navdeep;Kumar, Santosh;Raje, Manoj
通讯作者：
Raje, Manoj
影响因子：
--
作者：
Nagaoka M;Si-Tayeb K;Akaike T;Duncan SA
通讯作者：
Duncan SA
影响因子：
5.2
作者：
Bradley, Eric;Bieberich, Erhard;Mivechi, Nahid F.;Tangpisuthipongsa, Dantera;Wang, Guanghu
通讯作者：
Wang, Guanghu