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聚二甲基硅氧烷(PDMS)可调节CD38表达,吸收视黄酸并可能扰乱类视黄醇信号传导。

Polydimethylsiloxane (PDMS) modulates CD38 expression, absorbs retinoic acid and may perturb retinoid signalling.

作者信息

Futrega Kathryn, Yu Jianshi, Jones Jace W, Kane Maureen A, Lott William B, Atkinson Kerry, Doran Michael R

机构信息

Stem Cell Therapies Laboratory, Queensland University of Technology at the Translational Research Institute, 37 Kent Street Brisbane, QLD 4102, Australia.

Department of Pharmaceutical Sciences, University of Maryland School of Pharmacy, Baltimore, MD 21201, USA.

出版信息

Lab Chip. 2016 Apr 21;16(8):1473-83. doi: 10.1039/c6lc00269b.

Abstract

Polydimethylsiloxane (PDMS) is the most commonly used material in the manufacture of customized cell culture devices. While there is concern that uncured PDMS oligomers may leach into culture medium and/or hydrophobic molecules may be absorbed into PDMS structures, there is no consensus on how or if PDMS influences cell behaviour. We observed that human umbilical cord blood (CB)-derived CD34(+) cells expanded in standard culture medium on PDMS exhibit reduced CD38 surface expression, relative to cells cultured on tissue culture polystyrene (TCP). All-trans retinoic acid (ATRA) induces CD38 expression, and we reasoned that this hydrophobic molecule might be absorbed by PDMS. Through a series of experiments we demonstrated that ATRA-mediated CD38 expression was attenuated when cultures were maintained on PDMS. Medium pre-incubated on PDMS for extended durations resulted in a time-dependant reduction of ATRA in the medium and increasingly attenuated CD38 expression. This indicated a time-dependent absorption of ATRA into the PDMS. To better understand how PDMS might generally influence cell behaviour, Ingenuity Pathway Analysis (IPA) was used to identify potential upstream regulators. This analysis was performed for differentially expressed genes in primary cells including CD34(+) haematopoietic progenitor cells, mesenchymal stromal cells (MSC), and keratinocytes, and cell lines including prostate cancer epithelial cells (LNCaP), breast cancer epithelial cells (MCF-7), and myeloid leukaemia cells (KG1a). IPA predicted that the most likely common upstream regulator of perturbed pathways was ATRA. We demonstrate here that ATRA is absorbed by PDMS in a time-dependent manner and results in the concomitant reduced expression of CD38 on the cell surface of CB-derived CD34(+) cells.

摘要

聚二甲基硅氧烷(PDMS)是制造定制细胞培养装置时最常用的材料。尽管有人担心未固化的PDMS低聚物可能会渗入培养基,和/或疏水分子可能会被吸收到PDMS结构中,但关于PDMS如何影响或是否会影响细胞行为,目前尚无定论。我们观察到,与在组织培养聚苯乙烯(TCP)上培养的细胞相比,在PDMS上的标准培养基中培养的人脐带血(CB)来源的CD34(+)细胞,其CD38表面表达降低。全反式维甲酸(ATRA)可诱导CD38表达,我们推测这种疏水分子可能会被PDMS吸收。通过一系列实验,我们证明当细胞在PDMS上培养时,ATRA介导的CD38表达会减弱。在PDMS上预孵育较长时间的培养基会导致培养基中ATRA随时间减少,且CD38表达的减弱程度越来越大。这表明ATRA会随时间依赖性地被吸收到PDMS中。为了更好地理解PDMS通常如何影响细胞行为,我们使用了 Ingenuity 通路分析(IPA)来识别潜在的上游调节因子。对原代细胞(包括CD34(+)造血祖细胞、间充质基质细胞(MSC)和角质形成细胞)以及细胞系(包括前列腺癌上皮细胞(LNCaP)、乳腺癌上皮细胞(MCF-7)和髓系白血病细胞(KG1a))中差异表达的基因进行了该分析。IPA预测,受干扰通路最可能的共同上游调节因子是ATRA。我们在此证明,ATRA以时间依赖性方式被PDMS吸收,并导致CB来源的CD34(+)细胞表面CD38的表达随之降低。

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