Chung Bong Geun, Manbachi Amir, Khademhosseini Ali
Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA.
J Vis Exp. 2007(7):270. doi: 10.3791/270. Epub 2007 Aug 30.
We describe a microfluidic device with microgrooved patterns for studying cellular behavior. This microfluidic platform consists of a top fluidic channel and a bottom microgrooved substrate. To fabricate the microgrooved channels, a top poly(dimethylsiloxane) (PDMS) mold containing the impression of the microfluidic channels was aligned and bonded to a microgrooved substrate. Using this device, mouse fibroblast cells were immobilized and patterned within microgrooved substrates (25, 50, 75, and 100 microm wide). To study apoptosis in a microfluidic device, media containing hydrogen peroxide, Annexin V, and propidium iodide was perfused into the fluidic channel for 2 hours. We found that cells exposed to the oxidative stress became apoptotic. These apoptotic cells were confirmed by Annexin V that bound to phosphatidylserine at the outer leaflet of the plasma membrane during the apoptosis process. Using this microfluidic device with microgrooved patterns, the apoptosis process was observed in real-time and analyzed by using an inverted microscope containing an incubation chamber (37 degrees C, 5% CO(2)). Therefore, this microfluidic device incorporated with microgrooved substrates could be useful for studying the cellular behavior and performing high-throughput drug screening.
我们描述了一种带有微槽图案的微流控装置,用于研究细胞行为。这个微流控平台由一个顶部流体通道和一个底部微槽基板组成。为了制造微槽通道,将一个带有微流控通道印记的顶部聚二甲基硅氧烷(PDMS)模具与一个微槽基板对齐并键合。使用该装置,将小鼠成纤维细胞固定并在微槽基板(宽25、50、75和100微米)内进行图案化。为了研究微流控装置中的细胞凋亡,将含有过氧化氢、膜联蛋白V和碘化丙啶的培养基灌注到流体通道中2小时。我们发现暴露于氧化应激的细胞发生了凋亡。这些凋亡细胞通过膜联蛋白V得到证实,在凋亡过程中,膜联蛋白V与质膜外小叶上的磷脂酰丝氨酸结合。使用这种带有微槽图案的微流控装置,可以实时观察凋亡过程,并通过使用含有孵育室(37摄氏度,5%二氧化碳)的倒置显微镜进行分析。因此,这种结合了微槽基板的微流控装置可用于研究细胞行为和进行高通量药物筛选。