In the first study, researchers used a novel flow cytometry-based screen assessing 192 antibodies to cell surface markers to identify a cell surface signature that effectively distinguished neurons from hESC-derived neural stem cells (NSCs) and hESCs. Researchers used this cell surface signature in combination with flow cytometry to separate the undifferentiated cells from the differentiated cell types. This method reduces the variability of cell preparations from experiment-to-experiment and may help facilitate the development of assays requiring pure or consistent cell populations, such as genomics, transplantation experiments, in vitro drug testing and other in vitro assays where results may be confounded by unwanted cell types.
“Our study results help address a key challenge in the development of assays that will benefit from consistent, defined neural cell types,” said Christian Carson, Stem Cell Scientist, BD Biosciences and the study’s senior author. “In addition, these findings have implications beyond neural stem cells. They provide a framework of how to use flow cytometry analysis screens and sorting methods to quickly identify and isolate a variety of stem cells and their derivatives."
The second study demonstrates the successful application of the p160-Rho-associated coiled kinase (ROCK) inhibitor after cell sorting to improve cell survival in both feeder-dependent and feeder-independent growth conditions. Flow cytometry is a powerful tool used to effectively isolate and sort subpopulations of hESCs based on multiple markers, but due to the inherent sensitivity of hESCs, the recovery and survival of these cells can be low. However, the study found that by utilizing the ROCK inhibitor upon completion of cell sorting, researchers were able to improve recovery, enabling higher yield in experiments. Importantly, sorting and application of the ROCK inhibitor did not appear to negatively affect the cells. The sorted cells retain normal morphology, maintain a stable karyotype after many cell passages, and maintain their ability to differentiate into all three germ layers.
“Stem cell research is an increasingly complex and exhilarating area of science where researchers are looking to find answers to some of the most difficult to treat medical conditions,” says Jay Glasscock, President, Cell Analysis, BD Biosciences. “BD Biosciences is committed to working with the stem cell research community to help develop state-of-the-art tools that support and further enhance experiment capabilities.”
BD Biosciences offers a comprehensive portfolio of tools for stem cell researchers, spanning stem cell isolation, culture and analysis. BD Biosciences recently introduced five new stem cell research kits optimized for use on BD flow cytometry systems, including the latest launch of the BD™ Pluripotent Stem Cell Transcription Factor Analysis Kits. For more information about BD Biosciences’ newest stem cell research tools, please visit http://www.bdbiosciences.com/stemcellsource.
The study titled, “Isolation of a near-pure population of hESC-derived neurons using CD markers and fluorescence activated cell sorting (Abstract 2276)” was funded in collaboration with Lawrence S. Goldstein, Ph.D. and his laboratory at the University of California, San Diego.1 Its lead author was Shauna Yuan, M.D., a post-doctoral fellow.
About BD
BD is a leading global medical technology company that develops, manufactures and sells medical devices, instrument systems and reagents. The Company is dedicated to improving people's health throughout the world. BD is focused on improving drug delivery, enhancing the quality and speed of diagnosing infectious diseases and cancers, and advancing research, discovery and production of new drugs and vaccines. BD's capabilities are instrumental in combating many of the world's most pressing diseases. Founded in 1897 and headquartered in Franklin Lakes, New Jersey, BD employs approximately 28,000 people in approximately 50 countries throughout the world. The Company serves healthcare institutions, life science researchers, clinical laboratories, the pharmaceutical industry and the general public. For more information, please visit http://www.bd.com.
