Robotic Batch Somatic Cell Nuclear Transfer Based on Microfluidic Groove

Abstract

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.