Kalinec Gilda M, Park Channy, Thein Pru, Kalinec Federico
Head and Neck Surgery, David Geffen School of Medicine, University of California, Los Angeles.
Head and Neck Surgery, David Geffen School of Medicine, University of California, Los Angeles;
J Vis Exp. 2016 Sep 3(115):54425. doi: 10.3791/54425.
HEI-OC1 is one of the few mouse auditory cell lines available for research purposes. Originally proposed as an in vitro system for screening of ototoxic drugs, these cells have been used to investigate drug-activated apoptotic pathways, autophagy, senescence, mechanism of cell protection, inflammatory responses, cell differentiation, genetic and epigenetic effects of pharmacological drugs, effects of hypoxia, oxidative and endoplasmic reticulum stress, and expression of molecular channels and receptors. Among other several important markers of cochlear hair cells, HEI-OC1 cells endogenously express prestin, the paradigmatic motor protein of outer hair cells. Thus, they can be very useful to elucidate novel functional aspects of this important auditory protein. HEI-OC1 cells are very robust, and their culture usually does not present big complications. However, they require some special conditions such as avoiding the use of common anti-bacterial cocktails containing streptomycin or other antibiotics as well as incubation at 33 °C to stimulate cell proliferation and incubation at 39 °C to trigger cell differentiation. Here, we describe how to culture HEI-OC1 cells and how to use them in some typical assays, such as cell proliferation, viability, death, autophagy and senescence, as well as how to perform patch-clamp and non-linear capacitance measurements.
HEI-OC1是少数可用于研究目的的小鼠听觉细胞系之一。这些细胞最初被提议作为筛选耳毒性药物的体外系统，已被用于研究药物激活的凋亡途径、自噬、衰老、细胞保护机制、炎症反应、细胞分化、药理药物的遗传和表观遗传效应、缺氧、氧化和内质网应激的影响以及分子通道和受体的表达。在耳蜗毛细胞的其他几个重要标志物中，HEI-OC1细胞内源性表达prestin，这是外毛细胞的典型运动蛋白。因此，它们对于阐明这种重要听觉蛋白的新功能方面非常有用。HEI-OC1细胞非常强健，其培养通常不会出现大的并发症。然而，它们需要一些特殊条件，例如避免使用含有链霉素或其他抗生素的常见抗菌混合物，以及在33°C下孵育以刺激细胞增殖，在39°C下孵育以触发细胞分化。在这里，我们描述了如何培养HEI-OC1细胞以及如何在一些典型实验中使用它们，如细胞增殖、活力、死亡、自噬和衰老实验，以及如何进行膜片钳和非线性电容测量。
