Continuous Collection of Stem Cells from a Human Placenta Perfusion Co-Culture
Poster Mar 16, 2015
John J.S. Cadwell & James C. Hardy
The presence of smaller cells in the ECS harvests, OCT3/4 phenotype and the highly proliferative nature of this harvest when placed into flasks suggests the presence of very small embryonic-like stem cells. The hollow fiber cell culture environment is clearly different than flask culture. Three dimensional structures were formed on the surface of the fibers. Cytokine concentration and cell to cell interactions may play a role in their formation. When harvested cells were placed into flasks, the different cell culture environment induced these cells to differentiate into mesenchymal stem cells and generate spheroids. The recapitulation of the placental construct may represent a unique source for continuous culture and collection of stem cells. Hollow fiber bioreactors represent a different cell culture environment with resultant differences in cell phenotype from flask culture. These experiments demonstrate the capability of hollow fiber bioreactors to support continuous production of certain stem cell types in a single use system. Larger scale systems have the potential to produce stem cells on a continuous basis and meet the criteria for their bio-manufacturing in a single-use system on a clinical scale.
The nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) are closely related transcription factors that regulate the expression of phase I (cytochrome P450s), phase II metabolizing enzymes and transporter genes in response to xenobiotics, including prescription drugs.READ MORE
We found a distinct subpopulation of Tregs within BMSCs. Tregs and BMSCs in co-culture conferred neuroprotection that varied in a dose-dependent manner. Tregs minimized stem cell production of IL-6, a pro-inflammatory cytokine, and inhibited BMSC secretion of FGF-beta, a cytokine related to BMSC proliferation and differentiation. The ratio of Tregs found natively in BMSCs is optimally adapted to provide the maximum neuroprotective benefit of stem cell treatment after ischemic stroke.READ MORE
We utilized paired synthetic crRNAs coupled with our synthetic tracrRNA in cells transduced with lentiviral Cas9 to perform a functional knockout on hsa-miR-221. This three-part system (crRNA, tracrRNA and Cas9) has demonstrated efficient gene editing when used with only one guide RNA, but the goal was to use two crRNAs to remove the entire stem-loop.READ MORE