Robotic Batch Somatic Cell Nuclear Transfer Based on Microfluidic Groove

Somatic cell nuclear transfer (SCNT), which is an important procedure in cloning, has been conducted manually for decades. The operating efficiency drops sharply in batch SCNT because of the long-time observation under microscopy and the time-wasting traditional process. Though the operating time was reduced by robotic SCNT in previous studies, the traditional operating process was still used. In this article, we designed a new robotic batch SCNT process based on a microfluidic groove and two micropipettes in parallel. By using this new SCNT process, the operating area switching, objective lens conversing, and focusing on traditional SCNT process were eliminated, and oocyte localization was simplified, which saved much operating time. Experimental results showed that the new robotic batch process reduced about 50 s (41.7%) compared with the manual process (proposed 70 s versus manual 120 s). A success rate of 93.3% ( ${n} =30$ ) and a survival rate of 96.4% were achieved ( ${n}=28$ ), which were similar to manual process. The new robotic batch SCNT method demonstrated a high degree of efficiency and reproducibility. Note to Practitioners—This article presented a new robotic somatic cell nuclear transfer (SCNT) process. This new robotic SCNT process introduced a microfluidic groove for oocyte storage and two micropipettes for oocyte enucleation and oocyte injection, respectively. We save much operating time since the operating area switching, objective lens conversing, and focusing on traditional SCNT process were eliminated, and oocyte localization was simplified. Experimental results have demonstrated the efficiency and reproducibility of the new robotic SCNT process. This new robotic SCNT process has great potential for many other applications, e.g., ICSI, embryo microinjection, and cell biopsy. Commercialization of the proposed technology may lead to the improvement in SCNT industry. In current experiments, the somatic cells sometimes were injected at the same time, which led to the failure of the experiments. In the future, we will apply control algorithms to control the motion of multiple cells.

[1]  Xin Zhao,et al.  Pipelined batch-operation process of nuclear transplantation based on micro-manipulation system , 2016, 2016 IEEE International Conference on Robotics and Biomimetics (ROBIO).

[2]  Min Tan,et al.  Robotic Pick-And-Place of Multiple Embryos for Vitrification , 2017, IEEE Robotics and Automation Letters.

[3]  Tamio Tanikawa,et al.  Fluorescent monitoring using microfluidics chip and development of syringe pump for automation of enucleation to automate cloning , 2009, 2009 IEEE International Conference on Robotics and Automation.

[4]  I. Wilmut,et al.  Sheep cloned by nuclear transfer from a cultured cell line , 1996, Nature.

[5]  Mingzhu Sun,et al.  Robotic weighing for spherical cells based on falling speed detection , 2013, 2013 International Conference on Manipulation, Manufacturing and Measurement on the Nanoscale.

[6]  Zhiwei Zou,et al.  A Polymer Microfluidic Chip With Interdigitated Electrodes Arrays for Simultaneous Dielectrophoretic Manipulation and Impedimetric Detection of Microparticles , 2008, IEEE Sensors Journal.

[7]  Mingzhu Sun,et al.  Batch-operation process of nuclear transplantation based on global field of view , 2011, Proceedings of the 30th Chinese Control Conference.

[8]  Edward Grant,et al.  Blastocyst Microinjection Automation , 2009, IEEE Transactions on Information Technology in Biomedicine.

[9]  Win Tun Latt,et al.  Three-Dimensional Cell Rotation With Fluidic Flow-Controlled Cell Manipulating Device , 2016, IEEE/ASME Transactions on Mechatronics.

[10]  Yu Sun,et al.  A Fully Automated Robotic System for Microinjection of Zebrafish Embryos , 2007, PloS one.

[11]  Bradley J. Nelson,et al.  Biological Cell Injection Using an Autonomous MicroRobotic System , 2002, Int. J. Robotics Res..

[12]  Shaorong Xie,et al.  Robotic Adherent Cell Injection for Characterizing Cell–Cell Communication , 2015, IEEE Transactions on Biomedical Engineering.

[13]  Bijan Shirinzadeh,et al.  A novel cell weighing method based on the minimum immobilization pressure for biological applications , 2015 .

[14]  Xin Zhao,et al.  Robotic Cell Rotation Based on the Minimum Rotation Force , 2015, IEEE Transactions on Automation Science and Engineering.

[15]  Dong Sun,et al.  Achieving Automated Organelle Biopsy on Small Single Cells Using a Cell Surgery Robotic System , 2019, IEEE Transactions on Biomedical Engineering.

[16]  Johannes Courtial,et al.  Holographic assembly workstation for optical manipulation , 2008 .

[17]  Qingsong Xu,et al.  Micromachines for Biological Micromanipulation , 2018 .

[18]  S. Kishigami,et al.  Birth of cloned mice from vaginal smear cells after somatic cell nuclear transfer. , 2017, Theriogenology.

[19]  Clement Leung,et al.  Robotic ICSI (Intracytoplasmic Sperm Injection) , 2011, IEEE Transactions on Biomedical Engineering.

[20]  Yaowei Liu,et al.  Robotic Cell Rotation Based on Optimal Poking Direction , 2018, Micromachines.

[21]  Fumihito Arai,et al.  On-chip enucleation of an oocyte by untethered microrobots , 2014 .

[22]  L. E. Young,et al.  Somatic cell nuclear transfer , 2002, Nature.

[23]  Chen Feng,et al.  Autofocusing and Polar Body Detection in Automated Cell Manipulation , 2017, IEEE Transactions on Biomedical Engineering.

[24]  Dong Sun,et al.  A Force Control Approach to a Robot-assisted Cell Microinjection System , 2010, Int. J. Robotics Res..

[25]  G. Im,et al.  Dog cloning with in vivo matured oocytes obtained using electric chemiluminescence immunoassay-predicted ovulation method , 2017, PloS one.

[26]  Xin Zhao,et al.  Automated cell transportation for batch-cell manipulation , 2017, 2017 IEEE/RSJ International Conference on Intelligent Robots and Systems (IROS).

[27]  Dong Sun,et al.  A high-precision robot-aided single-cell biopsy system , 2017, 2017 IEEE International Conference on Robotics and Automation (ICRA).

[28]  Haibo Huang,et al.  Robotic Cell Injection System With Position and Force Control: Toward Automatic Batch Biomanipulation , 2009, IEEE Transactions on Robotics.

[29]  Y. Tsunoda,et al.  The Recent Progress on Nuclear Transfer in Mammals , 2000 .