High-Throughput Screens for Embryonic Stem Cells: Stress-Forced Potency-Stemness Loss Enables Toxicological Assays

2016 ◽  
pp. 279-312 ◽  
Author(s):  
Quanwen Li ◽  
Yu Yang ◽  
Erica Louden ◽  
Elizabeth E. Puscheck ◽  
Daniel A. Rappolee
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Embryonic Stem ◽  
High Throughput Screens

scholarly journals High-Throughput Screening Assay for the Identification of Compounds Regulating Self-Renewal and Differentiation in Human Embryonic Stem Cells

2008 ◽  
Vol 2 (6) ◽  
pp. 602-612 ◽  
Author(s):  
Sabrina C. Desbordes ◽  
Dimitris G. Placantonakis ◽  
Anthony Ciro ◽  
Nicholas D. Socci ◽  
Gabsang Lee ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Human Embryonic Stem Cells ◽  
High Throughput Screening ◽  
Embryonic Stem ◽  
Screening Assay ◽  
Self Renewal ◽  
High Throughput Screening Assay

scholarly journals Controlling Embryonic Stem Cell Growth and Differentiation by Automation

2012 ◽  
Vol 17 (9) ◽  
pp. 1171-1179 ◽  
Author(s):  
Michael P. Kowalski ◽  
Amy Yoder ◽  
Li Liu ◽  
Laura Pajak
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Drug Discovery ◽  
Embryonic Stem Cells ◽  
Embryonic Stem Cell ◽  
High Throughput ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Basic Research ◽  
Complex Biological Process

Despite significant use in basic research, embryonic stem cells have just begun to be used in the drug discovery process. Barriers to the adoption of embryonic stem cells in drug discovery include the difficulty in growing cells and inconsistent differentiation to the desired cellular phenotype. Embryonic stem cell cultures require consistent and frequent handling to maintain the cells in a pluripotent state. In addition, the preferred hanging drop method of embryoid body (EB) differentiation is not amenable to high-throughput methods, and suspension cultures of EBs show a high degree of variability. Murine embryonic stem cells passaged on an automated platform maintained ≥90% viability and pluripotency. We also developed a method of EB formation using 384-well microplates that form a single EB per well, with excellent uniformity across EBs. This format facilitated high-throughput differentiation and enabled screens to optimize directed differentiation into a desired cell type. Using this approach, we identified conditions that enhanced cardiomyocyte differentiation sevenfold. This optimized differentiation method showed excellent consistency for such a complex biological process. This automated approach to embryonic stem cell handling and differentiation can provide the high and consistent yields of differentiated cell types required for basic research, compound screens, and toxicity studies.

Precise Patterning of Mouse Embryonic Stem Cells on Glass Cover Slips for High-Throughput Analysis Using Laser Direct-Write

2011 ◽  
Author(s):  
Andrew D. Dias ◽  
Nathan R. Schiele ◽  
Brendan M. Carr ◽  
Nurazhani Abdul Raof ◽  
Yubing Xie ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Embryonic Stem ◽  
Direct Write ◽  
Glass Cover ◽  
Computer Aided ◽  
Petri Dishes ◽  
Laser Direct Write

Engineering a microenvironment where the growth substrate and distance between cells are controlled is highly desirable to understand how cellular interactions affect stem cell differentiation. Laser direct-write (LDW) allows rapid and precise placement of living cells via computer-aided design/computer-aided manufacturing (CAD/CAM) control. Application of this technique to study the effects of various stem cell microenvironments on differentiation requires a high-throughput experimental setup [1]. Recently, our lab has developed a gelatin-based LDW method for the precise patterning of sensitive cell types, such as mouse embryonic stem cells (mESCs), at a resolution of about 5 μm [2]. Although viable mESCs were successfully printed with maintained pluripotency, this technique required cells to be patterned onto polystyrene Petri dishes [2,3], which may limit high-throughput efficiency. Moreover, the use of polystyrene Petri dishes requires large quantities of culture medium and is not convenient for biological analysis of mESC differentiation. Therefore, the objective of this study was to adapt the LDW method, without altering its prior success, to transfer patterns of viable mESCs to glass cover slips. However, this adaptation to cover slips could not be achieved through simple downscaling due to the unique challenges of providing sufficient moisture for viable cell transfer while maintaining pattern registry on a cover slip. Once cells have been laser patterned, cover slips can then be moved to a 24-well plate so that separate sets of laser patterned cells can be analyzed in parallel for higher experimental throughput utilizing fewer resources to maintain the cells.

High-content screening of small compounds on human embryonic stem cells

2010 ◽  
Vol 38 (4) ◽  
pp. 1046-1050 ◽  
Author(s):  
Ivana Barbaric ◽  
Paul J. Gokhale ◽  
Peter W. Andrews
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Cell Biology ◽  
Es Cells ◽  
Embryonic Stem ◽  
High Content Screening ◽  
Cellular Models ◽  
Compound Libraries

Human ES (embryonic stem) cells and iPS (induced pluripotent stem) cells have been heralded as a source of differentiated cells that could be used in the treatment of degenerative diseases, such as Parkinson's disease or diabetes. Despite the great potential for their use in regenerative therapy, the challenge remains to understand the basic biology of these remarkable cells, in order to differentiate them into any functional cell type. Given the scale of the task, high-throughput screening of agents and culture conditions offers one way to accelerate these studies. The screening of small-compound libraries is particularly amenable to such high-throughput methods. Coupled with high-content screening technology that enables simultaneous assessment of multiple cellular features in an automated and quantitative way, this approach is proving powerful in identifying both small molecules as tools for manipulating stem cell fates and novel mechanisms of differentiation not previously associated with stem cell biology. Such screens performed on human ES cells also demonstrate the usefulness of human ES/iPS cells as cellular models for pharmacological testing of drug efficacy and toxicity, possibly a more imminent use of these cells than in regenerative medicine.

Array‐Based High‐Throughput Screening in Mouse Embryonic Stem Cells with sh RNAs

2013 ◽  
Vol 26 (1) ◽  
Author(s):  
Chia‐Hui Wang ◽  
Nianhan Ma ◽  
Yu‐Tsen Lin ◽  
Cheng‐Chung Wu ◽  
Hong‐Jin Wu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
High Throughput Screening ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cells

scholarly journals Efficient, high-throughput transfection of human embryonic stem cells

2010 ◽  
Vol 1 (3) ◽  
pp. 23 ◽  
Author(s):  
Jennifer C Moore ◽  
Kristin Atze ◽  
Percy L Yeung ◽  
Alana J Toro-Ramos ◽  
Cynthia Camarillo ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem

High throughput cryopreservation of cells by rapid freezing of sub-μl drops using inkjet printing – cryoprinting

Lab on a Chip ◽  
2015 ◽  
Vol 15 (17) ◽  
pp. 3503-3513 ◽  
Author(s):  
Rui Dou ◽  
Rachel E. Saunders ◽  
Lisa Mohamet ◽  
Christopher M. Ward ◽  
Brian Derby
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
High Throughput ◽  
Human Embryonic Stem Cells ◽  
Inkjet Printing ◽  
Embryonic Stem ◽  
Mouse Fibroblast ◽  
Rapid Freezing ◽  
Fibroblast Cells

We have successfully used inkjet printing to cryopreserve 3T3 mouse fibroblast cells and human neuroprogenitor cells (NPCs) derived from human embryonic stem cells (hESCs).

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