Selecting Monoclonal Cell Lineages from Somatic Reprogramming Using Robotic-Based Spatial-Restricting Structured Flow  

作　　者：Xueping Chen Ke Fan Jun Lu Sheng Zhang Jianhua Dong Jisheng Qin Weihua Fan Yan Wang Yiyuan Zhang Huo Peng Zhizhong Zhang Zhiyong Sun Chunlai Yu Yucui Xiong Yan Song Qingqing Ye Shiwen Mai Yuanhua Wang Qizheng Wang Fengxiang Zhang Xiaohui Wen Tiancheng Zhou Li Han Mian Long Guangjin Pan Julian F.Burke Xiao Zhang 

机构地区：[1]Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,People’s Republic of China [2]Institute of Mechanics,Chinese Academy of Sciences,Beijing 100190,People’s Republic of China [3]Institute of Electrical Engineering,Chinese Academy of Sciences,Beijing 100190,People’s Republic of China [4]CAS Key Laboratory of Regenerative Biology,Joint School of Life Sciences,Guangzhou Institutes of Biomedicine and Health,Chinese Academy of Sciences,Guangzhou 510530,People’s Republic of China,Guangzhou Medical University,Guangzhou 511436,People’s Republic of China [5]Biological Sciences,University of Southampton,University Road,Southampton SO171BJ,UK [6]School of Light Industry and Engineering,South China University of Technology,Guangzhou 510641,People’s Republic of China [7]University of Electronic Science and Technology of China,Chengdu 611731,People’s Republic of China

出　　处：《Research》2024年第4期442-457,共16页研究(英文)

基　　金：funded by the Key Project for Instrument Development Program-Ministry of Finance[Grant No.ZDYZ2012-3];Operational Support Project of Instrument Development-Chinese Academy of Sciences[Grant No.1187000169];Project of Automated Stem Cell Induction and Culture Instrument Development-Chinese Academy of Sciences[Grant No.1187000170];Scientific Instrument Development Program-Chinese Academy of Sciences[Grant Nos.ZDKYYQ20210006 and YJKYYQ20210042];Key Research Program of Chinese Academy of Sciences[Grant No.O2222001];Guangdong Basic and Applied Basic Research Foundation[Grant No.2022A1515110435];Guangzhou Basic and Applied Basic Research Project[Grant No.2023A04J0107];Project funded by China Postdoctoral Science Foundation[Grant No.2021M693192];Guangdong International Scientific Research Cooperation Project[Grant No.2022A0505050037];Science and Technology Planning Project of Guangdong Province[Grant No.2023B1212060050].

摘　　要：Somatic cell reprogramming generates induced pluripotent stem cells(iPSCs),which serve as a crucial source of seed cells for personalized disease modeling and treatment in regenerative medicine.However,the process of reprogramming often causes substantial lineage manipulations,thereby increasing cellular heterogeneity.As a consequence,the process of harvesting monoclonal iPSCs is labor-intensive and leads to decreased reproducibility.Here,we report the first in-house developed robotic platform that uses a pin-tip-based micro-structure to manipulate radial shear flow for automated monoclonal iPSC colony selection(~1 s)in a non-invasive and label-free manner,which includes tasks for somatic cell reprogramming culturing,medium changes;time-lapse-based high-content imaging;and iPSCs monoclonal colony detection,selection,and expansion.Throughput-wise,this automated robotic system can perform approximately 24 somatic cell reprogramming tasks within 50 days in parallel via a scheduling program.Moreover,thanks to a dual flow-based iPSC selection process,the purity of iPSCs was enhanced,while simultaneously eliminating the need for single-cell subcloning.These iPSCs generated via the dual processing robotic approach demonstrated a purity 3.7 times greater than that of the conventional manual methods.In addition,the automatically produced human iPSCs exhibited typical pluripotent transcriptional profiles,differentiation potential,and karyotypes.In conclusion,this robotic method could offer a promising solution for the automated isolation or purification of lineage-specific cells derived from iPSCs,thereby accelerating the development of personalized medicines.

关 键 词：PROGRAMMING thereby PURITY 

分 类 号：R32[医药卫生—人体解剖和组织胚胎学]