摘要
细胞从培养皿表层脱壁是贴壁细胞传代的必须操作,目前,胰蛋白酶消化法是细胞脱壁的主要方法,但该方法操作复杂,难以实现自动化细胞培养。此外,胰蛋白酶还会对细胞蛋白造成不可逆的伤害,造成细胞产量下降及自修复时间长等问题。超声脱壁技术使用物理手段完成细胞脱壁,可以避免损伤细胞蛋白,实现脱壁过程的自动化。该文研究了一种超声脱壁技术,采用基于亮场显微镜的细胞融合度检测方法,对超声脱壁技术的脱壁效果进行评估,以人脐带间充质干细胞为目标细胞,研究了贴壁细胞超声脱壁技术的关键控制参数。实验结果显示,当超声换能器输入电压为255 V、超声频率为159~161 kHz扫频、超声时间为2 min时,将细胞与超声换能器距离调至3 mm,使用12 mL生理盐水作为震荡液体,可以使T75瓶中细胞脱壁面积达到95%,细胞成活率达到70%。
Detaching cells from the surface of the flask is a necessary operation for the adherent cell passage. Trypsinization is currently the primary method for cell detachment. However, this method is complicated to automated cell culture. Trypsin can cause irreversible damage to cellular proteins,decreasing of cell yield and long self-repair time. Ultrasonic detachment technology uses physical means to complete cell detachment operations, which can avoid damage to cellular proteins and make the realization of automated detachment easier. In this paper, we have developed a device for ultrasonic detachment.Human umbilical cord mesenchymal stem cells were used to study ultrasonic detachment technology control parameters. The experiment results showed that, the optimum cell detachment was achieved(95%detachment rate and 70% survival rate) in 12 mL 5%-saline with 255 V and 159~161 kHz sweeping input signal, 2 min stimulation period, and 3 mm space between flask surface and transducer.
作者
陈晓彤
王泽南
魏军
胡颖
CHEN Xiaotong;WANG Zenan;WEI Jun;HU Ying(School of Automation and Electrical Engineering,University of Jinan,Jinan 250024,China;Shenzhen Institute of Advanced Technology,Chinese Academy of Sciences,Shenzhen 518055,China)
出处
《集成技术》
2022年第5期1-11,共11页
Journal of Integration Technology
基金
广东省重点领域研发计划项目(2022B1111010003)
国家自然科学基金项目(62050410349)
深圳市重点基础研究项目(JCYJ20210324115601004)。
关键词
超声脱壁技术
自动化控制
细胞脱壁
细胞实验
ultrasonic detachment technology
automatic control
cell detachment
cell experiment
