Schrader S, Kremling C, Klinger M, Laqua H, Geerling G
Department of Ophthalmology, University of Lübeck, Lübeck, Germany.
Br J Ophthalmol. 2009 Aug;93(8):1121-5. doi: 10.1136/bjo.2008.137927. Epub 2009 May 4.
A rotary cell-culture system (RCCS) allows the creation of a microgravity environment of low shear force, high-mass transfer and three-dimensional cell culture of various cell types. The aim of the study was to evaluate the growth pattern and the secretory function of rabbit lacrimal gland acinar cells in a microgravity environment using an RCCS.
Lacrimal gland acinar cells from male New Zealand White rabbits were isolated and cultured in an RCCS up to 28 days. Cells were analysed by light and electron microscopy, and apoptosis was assessed by the TUNEL assay at days 7, 14, 21 and 28. Secretory function was tested by measuring the beta-hexosaminidase activity.
After 7 days of culture, spheroidal aggregates were found inside the RCCS. The spheroids consisted of acinus-like cell conglomerates. Apoptotic centres inside the spheroids were observed at all time points by means of the TUNEL assay. Evaluation of the secretory function revealed beta-hexosaminidase release after carbachol stimulation which decreased over the culture period.
A simulated microgravity environment promotes the development of three-dimensional cell spheroids containing viable acinar cells up to 28 days. Due to the evolving central apoptosis, it is unlikely that such simple three-dimensional cell communities can serve as tissue equivalents for clinical transplantation, but they promise opportunities for further applications in basic and applied cell research on lacrimal gland cells.
旋转细胞培养系统（RCCS）能够创建一个低剪切力、高传质且适用于多种细胞类型进行三维细胞培养的微重力环境。本研究旨在使用RCCS评估兔泪腺腺泡细胞在微重力环境下的生长模式和分泌功能。
从雄性新西兰白兔分离泪腺腺泡细胞，并在RCCS中培养长达28天。通过光学显微镜和电子显微镜对细胞进行分析，并在第7、14、21和28天通过TUNEL检测评估细胞凋亡情况。通过测量β-己糖胺酶活性来测试分泌功能。
培养7天后，在RCCS内部发现了球状聚集体。这些球体由腺泡样细胞团组成。通过TUNEL检测在所有时间点均观察到球体内的凋亡中心。对分泌功能的评估显示，卡巴胆碱刺激后β-己糖胺酶释放，且在培养期间有所下降。
模拟微重力环境可促进含有活腺泡细胞的三维细胞球体的形成，长达28天。由于中央凋亡的演变，这种简单的三维细胞群落不太可能作为临床移植的组织等效物，但它们为泪腺细胞的基础和应用细胞研究提供了进一步应用的机会。
