Gong Yong-Kuan, Luo Li, Petit Alain, Zukor David J, Huk Olga L, Antoniou John, Winnik Françoise M, Mwale Fackson
Department of Chemistry and Faculty of Pharmacy, University of Montreal, CP 6128, Succursale Centre Ville, Montreal, Quebec, H3C 3J7, Canada.
J Biomed Mater Res A. 2005 Jan 1;72(1):1-9. doi: 10.1002/jbm.a.30135.
A new type of amphiphilic phosphorylcholine (PC) polymer was used in this work to develop a cell culture surface that allows the attachment of U937 macrophages. The PC polymer was a random copolymer of N-isopropylacrylamide (45%), N-(phosphorylcholine)-N'-(ethylenedioxy-bis(ethyl)) acrylamide (41%), and the hydrophobic monomer N-(n-octadecyl) acrylamide (14%). Polypropylene (PP) films (1 cm2) were coated with the polymer solution by immersion. U937 macrophage suspensions were applied on PC polymer-coated surfaces and incubated for up to 72 h at 37 degrees C. While U937 cells did not adhere to PP, ammonia, nitrogen, or oxygen plasma-treated surfaces, they attached rapidly on PC-coated surfaces (< 1 h), proliferated, and stayed attached to the modified surface for at least 72 h, suggesting that unique features of the PC polymer, and the U937 macrophages, are responsible for the attachment of these cells. We compared the effect of Co2+ and Cr3+ ions on the expression of bone-resorbing cytokines (TNF-alpha, IL-6, IL-1beta) in U937 macrophages cultured on PC-coated surfaces to the response of U937 macrophages in suspension. Cytokine gene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). Addition of Co2+ and Cr3+ ions led to a significant increased expression of TNF-alpha in both cultured and suspension cells. On the other hand, Co2+ and Cr3+ ions had a weak stimulatory effect or no effect on IL-1beta and IL-6, respectively, in both cultured and suspension cells. In conclusion, the use of PC polymer-modified surfaces might offer promising new opportunities for the culture of human U937 cells and may also point to the mechanism by which macrophages interact with lipid bilayers of biological membranes.
在本研究中，一种新型两亲性磷酰胆碱（PC）聚合物被用于开发一种可使U937巨噬细胞附着的细胞培养表面。该PC聚合物是N - 异丙基丙烯酰胺（45%）、N -（磷酰胆碱）- N'-（乙二氧基 - 双（乙基））丙烯酰胺（41%）和疏水性单体N -（正十八烷基）丙烯酰胺（14%）的无规共聚物。通过浸泡法将聚合物溶液涂覆在聚丙烯（PP）薄膜（1平方厘米）上。将U937巨噬细胞悬液接种在PC聚合物涂覆的表面上，并在37℃下孵育长达72小时。虽然U937细胞不附着于PP、氨、氮或氧等离子体处理过的表面，但它们能迅速附着在PC涂覆的表面上（<1小时），进行增殖，并在修饰表面上至少保持附着72小时，这表明PC聚合物的独特特性以及U937巨噬细胞共同促成了这些细胞的附着。我们比较了Co2 +和Cr3 +离子对在PC涂覆表面上培养的U937巨噬细胞中骨吸收细胞因子（TNF -α、IL - 6、IL - 1β）表达的影响与悬浮培养的U937巨噬细胞的反应。通过逆转录聚合酶链反应（RT - PCR）分析细胞因子基因表达。添加Co2 +和Cr3 +离子导致培养细胞和悬浮细胞中TNF -α的表达均显著增加。另一方面，Co2 +和Cr3 +离子对培养细胞和悬浮细胞中的IL - 1β和IL - 6分别具有微弱的刺激作用或无作用。总之，使用PC聚合物修饰的表面可能为人类U937细胞的培养提供有前景的新机会，并且还可能揭示巨噬细胞与生物膜脂质双层相互作用的机制。
