Maintenance of Pluripotency in Human Embryonic Stem Cells Cultured on a Synthetic Substrate in Conditioned Medium
Maintenance of Pluripotency in Human Embryonic Stem Cells Cultured on a Synthetic Substrate in Conditioned Medium
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DOI：
10.1002/bit.22520
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发表时间：
2010-01-01
影响因子：
3.8
通讯作者：
Denning, Chris
中科院分区：
文献类型：
--
作者：
Mahlstedt, Magdalena M.;Anderson, David;Denning, Chris
实现人类胚胎干细胞（hESC）的潜在临床和工业应用受到昂贵，不稳定或不确定的生长基质的限制。在这里，我们证明了在条件培养基中的氧等离子体蚀刻的组织培养聚苯乙烯（PE-TCPS）上的hESC系HUES 7和N 0 TT 1的胰蛋白酶传代与多能性相容。该合成培养表面在室温下稳定至少一年，并且通过将聚苯乙烯基底置于射频氧等离子体发生器中5分钟而容易地制备。通过X射线光电子能谱（XPS）和飞行时间二次离子质谱证实了通过等离子体蚀刻的聚苯乙烯表面化学的改性（ToF-SIMS），其鉴定作为处理结果的元素和分子变化。在PE-TCPS上培养的hESC的多能性通过连续传代期间的一致增殖、干细胞标志物（OCT 4、TRA 1 -60和SSEA-4）的表达、稳定的核型和体外多胚层分化（包括向对细胞增殖反应的心肌细胞的分化）来衡量。生成具有成本效益的，易于处理的合成的，确定的，稳定的表面hESC培养将加快干细胞在生物医学应用中的使用。Biotechnol. Bioeng. 2010;105：130-140. (C)2009 Wiley Periodicals，Inc.
Realizing the potential clinical and industrial applications of human embryonic stem cells (hESCs) is limited by the need for costly, labile, or undefined growth substrates. Here we demonstrate that trypsin passaging of the hESC lines, HUES7 and NOTT1, on oxygen plasma etched tissue culture polystyrene (PE-TCPS) in conditioned medium is compatible with pluripotency. This synthetic culture surface is stable at room temperature for at least a year and is readily prepared by placing polystyrene substrates in a radio frequency oxygen plasma generator for 5 min. Modification of the polystyrene surface chemistry by plasma etching was confirmed by X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS), which identified elemental and molecular changes as a result of the treatment. Pluripotency of hESCs cultured on PE-TCPS was gauged by consistent proliferation during serial passage, expression of stem cell markers (OCT4, TRA1-60, and SSEA-4), stable karyotype and multi-germlayer differentiation in vitro, including to pharmacologically responsive cardiomyocytes. Generation of cost-effective, easy-to-handle synthetic, defined, stable surfaces for hESC culture will expedite stem cell use in biomedical applications. Biotechnol. Bioeng. 2010;105: 130-140. (C) 2009 Wiley Periodicals, Inc.