Full Length Research Paper
Silin Sa1, Diep T. Nguyen3, Jonathan D. Pegan2, Michelle Khine3 and Kara E. McCloskey1,2* 1Graduate Group in
1Biological Engineering and Small-scale Technologies, University of California, Merced, USA.
2School of Engineering, University of California, Merced, USA. 3Department of Biomedical Engineering, University of California, Irvine, USA.
The differentiation of embryonic stem cells (ESC) into tissue-specific cells utilizes either monolayer cultures or three-dimensional cell aggregates called embryoid bodies (EB). However, the generation of a large number of EB of controlled sizes can be challenging and labor intensive. Our laboratories have developed a simple, robust, ultra-rapid, and inexpensive design of Honeycomb Microwells for generation of EB. Here, we compare EB generated using (1) Honeycomb Microwells, (2) the commercially available AggreWell™400, and (3) the more traditional Hanging Drop method. We compared the efficiency, viability, quality, and control of EB sizes. Results indicate that the Honeycomb Microwell and AggreWell™400 efficiently generate small EB at approximately 500 cells per EB. However, the cone-bottomed AggreWell plate generates cone-shaped EB at 1000-2000 cells per EB. Moreover, the cone-shape correlates with a reduction in the formation of the primitive endoderm GATA-4+ cells (1% compared with 6-8% in spherical EB), but does not significantly affect mesoderm or ectoderm development. We conclude that the non-spherical EB shape correlates with a reduction in the development of primitive endoderm, and that use of these AggreWell plates should be avoided in deriving endoderm tissue products. Key words: Embryonic stem cells, embryoid bodies, Microwell, AggreWell, hanging drop, primitive endoderm, GATA-4.
|APA||(2012). Round-bottomed Honeycomb Microwells: Embryoid body shape correlates with stem cell fate. Journal of Developmental Biology and Tissue Engineering, 5(2), 12-22.|
|Chicago||Silin Sa, Diep T. Nguyen, Jonathan D. Pegan, Michelle Khine and Kara E. McCloskey, Graduate Group in . "Round-bottomed Honeycomb Microwells: Embryoid body shape correlates with stem cell fate." Journal of Developmental Biology and Tissue Engineering 5, no. 2 (2012): 12-22.|
|MLA||Silin Sa, et al. "Round-bottomed Honeycomb Microwells: Embryoid body shape correlates with stem cell fate." Journal of Developmental Biology and Tissue Engineering 5.2 (2012): 12-22.|