My work concerns the improvement of an in vitro protocol to obtain a large scale production of red blood cells for transfusion, starting from PBMC. Particularly, my project focuses on the purification of reticulocytes obtained through cell culture. It has been shown that at the end of the differentiation process it is possible to find in our culture system three different cell populations: nucleated red blood cells (nRBC) cells which still have to go through the enucleation process, pyrenocytes (the nuclei of the cells) and reticulocytes or enucleated red blood cells (eRBC) which are our cells of interest.
Generally, in vivo, pyrenocytes are depleted through macrophages, bus since in our cultures there are any macrophages, it becomes necessary to find a way to remove this cells from our final product.
The idea of the project comes from this background. We would like to identify a protein (or even more than one) only expressed on the pyrenocytes and nRBC surface and to build an aptamers that can bind the target protein in a very specific way. The aptamer then will be associated with a selection structure, such as a filter or a column in order to purificate eRBC from pyrenocytes and nRBC that will bind to the aptamer.