Comparison of various longer term cell labeling methods on embryonic stem cells

• Main Author: Chong, Rui Qi
• Other Authors: Wang Dongan
• Format: Final Year Project
• Language: English
• Published: 2016
• Subjects: DRNTU::Engineering
• Online Access: http://hdl.handle.net/10356/68338
• Institution: Nanyang Technological University

Mouse embryonic stem cells are widely used to investigate the early embryonic development and differentiation of these cells, and are therefore widely used in applications such as biomedical research and regenerative medicine nowadays. However, these investigations are limited without a proper cell labeling method. Hence, this paper aims to investigate and compare the various longer term cell labeling methods on the monolayer of the embryonic stem cells, so that the biodistribution of cells and other functional attributes could be studied thoroughly. The 3 markers to be compared are PLGA nanoparticle based sensor platform, green fluorescent proteins and DiO cell labeling solution. The PLGA nanoparticle based sensor platform is synthesized by encapsulating Calcein Acetomethoxy (CAM) inside the PLGA nanoparticle to track and monitor the cells. CAM emits fluorescence once there is uptake by the cells. DiO and GFP plasmid are widely used cell labeling markers. Labeling efficiency and stability using the three methods was assessed after day 1, day 3 and day 7 of cell labeling. Proliferation rate and pluripotency of the cells , using pluripotency markers were also examined, so as to give a more complete comparison of the 3 types of markers used to label cells. Lastly, confocal microscopy is used to visualize the EB formation process. According to the results, it is concluded that cells labeled with PLGA nanoparticles are labeled efficiently, and are able to report cell status, and does not affect the formation of embryoid bodies, and hence could be used as a novel labeling method.