Surface modification of PHBV nanofiber mats for rapid cell cultivation and harvesting
Surface modification of PHBV nanofiber mats for rapid cell cultivation and harvesting
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DOI：
10.1080/09205063.2017.1414481
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发表时间：
2018-01-01
影响因子：
3.6
通讯作者：
Kwon, Oh Hyeong
中科院分区：
文献类型：
--
作者：
Ko, Young-Gwang;Kim, Young-Jin;Kwon, Oh Hyeong
为了保持特定组织的原有功能以用于治疗性组织工程，需要一种先进的细胞培养表面来重复细胞增殖。我们设计了一种新型的细胞增殖和快速收获表面，将非织造纳米纤维垫和一种温敏性聚合物结合在一起。采用静电纺丝技术制备了聚(3-羟基丁酸酯-3-羟基戊酸酯)纳米纤维膜。采用电子束辐照的方法在PHBV型纳米纤维毡上接枝了聚N-异丙基丙烯酰胺(PNIPAM)热响应层。用扫描电子显微镜测定了PNIPAM接枝PHBV型纳米纤维的平均直径。采用ATR-FTIR和ESCA对PNIPAM接枝到PHBV型纳米纤维表面进行了表征。测量了垫子上的水接触角对温度变化的响应。将人脂肪干细胞(ADSCs)培养在PNIPAM接枝的PHBV型纳米纤维垫上，以研究重复传代过程中细胞的增殖、收获和功能。用荧光激活细胞分类器(FACS)比较低温处理和胰酶-EDTA从每个表面分离的ADSCs表达的干细胞膜特异性标志物，如CD-13 PE、CD-29 PE和CD-90 FITC。利用温度响应型PHBV型纳米纤维垫通过低温处理大量培养和完整获取干细胞是一种在再生医学和干细胞治疗中应用的有前途的技术。
To maintain the original function of a specific tissue for therapeutic tissue engineering, an advanced cell culture surface for repeat cell proliferation is necessary. We designed a novel cell proliferation and rapid harvesting surface by combining nonwoven nanofiber mat and a thermo-responsive polymer. Nanofibrous poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) mats were fabricated by the electrospinning technique. A poly(N-isopropylacrylamide) (PNIPAM) thermo-responsive layer was grafted on the PHBV nanofiber mat by electron beam irradiation. The average diameter of the PNIPAM-grafted PHBV nanofibers was determined by SEM. ATR-FTIR and ESCA were used to confirm the grafting of PNIPAM onto the PHBV nanofiber surface. Water contact angles on the mats were measured in response to temperature changes. Human adipose-derived stem cells (ADSCs) were cultured on the PNIPAM-grafted PHBV nanofiber mat to investigate cell proliferation, harvesting, and functionality during repeat subculture. Detached ADSCs from each surface by low temperature treatment and trypsin-EDTA were compared by a fluorescence-activated cell sorter (FACS) using expression of stem cell membrane-specific markers such as CD-13 PE, CD-29 PE, and CD-90 FITC. The mass cultivation and intact harvesting of stem cells by low temperature treatment using a thermo-responsive PHBV nanofiber mat is a promising technique for use in regenerative medicine and stem cell therapy.