Simulating Embryonic Development in Vitro Using Embryoid Bodies

Overview

During the developmental stages of mammalian embryos, the inner cell mass (ICM) gives rise to all embryonic tissues. The ICM, when adapted to in vitro culture, becomes embryonic stem cells (ESCs). These pluripotent ESCs are capable of differentiating into all three germ-layers. The purpose of the investigation is to reinitiate embryonic development in vitro using a mouse reporter ESC line and to observe self-organization of the germ- layers. The reporter ESCs express yellow fluorescent protein when the mesoderm (one of the germ-layers) specific gene, Brachyury, is activated. ESCs are often cultured into 3D cell aggregates which mimic the traits of early embryonic development, called embryoid bodies (EBs). Here, two methods were used to generate EBs- the hanging drop culture and Aggrewell 400 plates. In hanging drop culture, ESC containing droplets were dispensed onto the underside of a lid of a tissue culture plate and flipped over so that the droplets were suspended over water to prevent evaporation. After 4 days, the EBs were collected and counted. In comparison, for the Aggrewell 400 plates, an anti-adherence solution was coated and rinsed off before adding cells to each one of the 24 wells. After 4 days, EBs were counted. EBs from both methods were imaged using an inverted epifluorescence widefield microscope and the images were processed using the ImageJ software. Based on this investigation, the hanging drop culture method generated more spherical EBs and allowed for mesoderm germ- layer to be organized “in the middle” of the EBs compared to the Aggrewell 400 plates. Integrating 3D Printing into Titanium Dioxide Nanotube Fabrication Ryan He University of Illinois Laboratory High School, Urbana, IL Supervisor: Dr. Hongshan He Titanium dioxide nanotubes (TiO2 NTs) have numerous biomedical, photocatalytic, and optical properties. However, a noticeable trend in the literature is the absence of electrode spacing parameters. Fortunately, this confounding variable can be eliminated with the usage of 3D printing and CAD. By integrating materials chemistry and rapid prototyping, unspecified parameters can be standardized by means of a 3D printed anodization guide. Herein, several nanotube growth guides were fabricated and tested within an electrolytic cell to evaluate their effectiveness and region-dependent growth trends. Observational characterization demonstrates that TiO2 NTs were observed to grow longer and with wider diameters at areas closer to the opposite electrode while others grew shorter and had smaller diameters at farther areas. These data informed a transitional approach to implementing the templates in the direct synthesis of TiO2 nanotubes on conductive FTO glass. The results from this testing show novel growth patterns and allow for future improvements for facile syntheses of TiO2 NTs on various surfaces. We conclude that this demonstrates the versatility and usability of 3D printing in the laboratory as a response to certain reaction conditions.

Competition history

• JSHS 2023 Category not listed

Resources

• National JSHS abstract book ↗

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