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封闭圆形微通道的简易台式制造为前列腺上皮细胞的生长提供了三维受限结构。

Facile bench-top fabrication of enclosed circular microchannels provides 3D confined structure for growth of prostate epithelial cells.

作者信息

Dolega Monika E, Wagh Jayesh, Gerbaud Sophie, Kermarrec Frederique, Alcaraz Jean-Pierre, Martin Donald K, Gidrol Xavier, Picollet-D'hahan Nathalie

机构信息

Univ. Grenoble Alpes, iRTSV-BGE, Grenoble, France; CEA, iRTSV-BGE, Grenoble, France; INSERM, BGE, Grenoble, France.

UJF-Grenoble 1, CNRS, TIMC-IMAG UMR 5525 (SyNaBi), Grenoble, France.

出版信息

PLoS One. 2014 Jun 19;9(6):e99416. doi: 10.1371/journal.pone.0099416. eCollection 2014.

Abstract

We present a simple bench-top method to fabricate enclosed circular channels for biological experiments. Fabricating the channels takes less than 2 hours by using glass capillaries of various diameters (from 100 µm up to 400 µm) as a mould in PDMS. The inner surface of microchannels prepared in this way was coated with a thin membrane of either Matrigel or a layer-by-layer polyelectrolyte to control cellular adhesion. The microchannels were then used as scaffolds for 3D-confined epithelial cell culture. To show that our device can be used with several epithelial cell types from exocrine glandular tissues, we performed our biological studies on adherent epithelial prostate cells (non-malignant RWPE-1 and invasive PC3) and also on breast (non-malignant MCF10A) cells We observed that in static conditions cells adhere and proliferate to form a confluent layer in channels of 150 µm in diameter and larger, whereas cellular viability decreases with decreasing diameter of the channel. Matrigel and PSS (poly (sodium 4-styrenesulphonate)) promote cell adhesion, whereas the cell proliferation rate was reduced on the PAH (poly (allylamine hydrochloride))-terminated surface. Moreover infusing channels with a continuous flow did not induce any cellular detachment. Our system is designed to simply grow cells in a microchannel structure and could be easily fabricated in any biological laboratory. It offers opportunities to grow epithelial cells that support the formation of a light. This system could be eventually used, for example, to collect cellular secretions, or study cell responses to graduated hypoxia conditions, to chemicals (drugs, siRNA, …) and/or physiological shear stress.

摘要

我们提出了一种简单的台式方法来制造用于生物实验的封闭圆形通道。通过使用各种直径(从100 µm到400 µm)的玻璃毛细管作为聚二甲基硅氧烷(PDMS)中的模具来制造通道,耗时不到2小时。以这种方式制备的微通道内表面涂有一层基质胶薄膜或层层聚电解质,以控制细胞粘附。然后将这些微通道用作三维受限上皮细胞培养的支架。为了表明我们的装置可用于来自外分泌腺组织的几种上皮细胞类型,我们对贴壁上皮前列腺细胞(非恶性RWPE-1和侵袭性PC3)以及乳腺(非恶性MCF10A)细胞进行了生物学研究。我们观察到,在静态条件下,细胞在直径为150 µm及更大的通道中粘附并增殖形成汇合层,而细胞活力随着通道直径的减小而降低。基质胶和聚苯乙烯磺酸钠(PSS)促进细胞粘附,而在聚烯丙胺盐酸盐(PAH)封端的表面上细胞增殖速率降低。此外,向通道中注入连续流不会导致任何细胞脱离。我们的系统旨在简单地在微通道结构中培养细胞,并且可以在任何生物实验室中轻松制造。它为培养支持光形成的上皮细胞提供了机会。例如,该系统最终可用于收集细胞分泌物,或研究细胞对渐进性缺氧条件、化学物质(药物、小干扰RNA等)和/或生理剪切应力的反应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/43f0/4063722/f7488fe37b61/pone.0099416.g001.jpg

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