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

机译:聚二甲基硅氧烷(PDMS)调节CD38表达,吸收视黄酸并可能干扰类视色素信号

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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.
机译:聚二甲基硅氧烷(PDMS)是定制细胞培养设备制造中最常用的材料。尽管担心未固化的PDMS低聚物可能会浸入培养基中和/或疏水性分子可能被PDMS结构吸收,但关于PDMS如何或是否影响细胞行为尚无共识。我们观察到,相对于组织培养聚苯乙烯(TCP)上培养的细胞,在PDMS的标准培养基中扩增的人脐带血(CB)衍生的CD34(+)细胞显示出降低的CD38表面表达。全反式视黄酸(ATRA)诱导CD38表达,我们认为该疏水分子可能被PDMS吸收。通过一系列实验,我们证明当将培养物保持在PDMS上时,ATRA介导的CD38表达减弱。在PDMS上预孵育的培养基延长的时间导致培养基中ATRA的时间依赖性降低,并逐渐减弱CD38的表达。这表明ATRA被PDMS吸收与时间有关。为了更好地了解PDMS一般如何影响细胞行为,使用了Ingenuity Pathway Analysis(IPA)来识别潜在的上游调节剂。对包括CD34 +造血祖细胞,间充质基质细胞(MSC)和角质形成细胞在内的原代细胞以及包括前列腺癌上皮细胞(LNCaP),乳腺癌上皮细胞(MCF-7)和髓样白血病细胞(KG1a)。 IPA预测,最可能的扰动途径常见上游调节剂是ATRA。我们在这里证明,ATRA被PDMS以时间依赖的方式吸收,并导致CB衍生的CD34(+)细胞表面上CD38的表达减少。

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