Development of a Novel Xeno-free Medium for Feeder-free Culture of Human Stem Cells
Poster Mar 20, 2015
Annand R; Okuda Y; Inamura M
Culture of human ES cells and iPS cells has attracted a lot of interest due to the applications of stem cells in both drug screening as well as regenerative medicine.
Most researchers co-cultivate human ES or iPS cells on mouse derived MEF feeder cells. However, presence of the feeder cells could affect the application of the resulting stem cell to other areas of research. Moreover, the current conventional culture medium may contain animal-derived serum; this may increase the risk transspecies infection from implantation of stem cells or stem-cell derived materials. Such issues could cause a setback in the clinical application of stem cell research. In order to solve such issues, we have developed a new culture medium, ReproXFTM, which does not contain any animal-derived components. Culture with ReproXFTM allows researchers to cultivate human iPS and ES cells under feeder-free conditions without compromising the quality.
Here, we demonstrate that human iPS cells cultivated using ReproXFTM showed alkaline phosphatase activity. Furthermore, these iPS cells showed strong expression of the pluripotency markers OCT3/4, NANOG, SSEA-1, TRA1-60 and TRA1-80 by both
immunostaining and flow cytometry. Also, from the result of immunostaining, we have confirmed that the karyotype of these iPS cells is normal. In addition, we also confirmed that human iPS cells cultivated in ReproXFTM possess the ability to differentiate into neurons and cardiomyocytes. Taken together, these data show that ReproXFTM medium not only allows human iPS cells to remain in the undifferentiated, pluripotent state, but also to retain the ability to
differentiate under standard conditions.
In conclusion, we believe that ReproXFTM medium will allow researchers to produce large amounts of high quality of human ES or iPS cells for use in regenerative medicine and basic research.
Multiplexing cell-based assays is possible using 3D culture models that are larger and more complex than monolayers
Real-time detection methods to measure live or dead cells provide much flexibility for multiplexing
All multiplexed assay combinations should be verified using appropriate controls for each 3D cell culture model.
Basic fibroblast growth factor (bFGF) is widely used in vitro for the maintenance and stimulation of a variety of cells. However, use of native bFGF in cell biology is limited by the fact that bFGF rapidly degrades at physiological temperatures. We have addressed this problem with an engineered form of bFGF, named Heat Stable bFGF (HS bFGF), which is stable at 37 degrees Celsius.READ MORE