Characterization of Exosomes from Alveolar Macrophages and CFTR Expression
Exosomes are 30-100nm sized vesicles that play a role in cellular communication due to their ability to transfer proteins and nucleic acids. Characterization of the protein and nucleic acids that are transferred from each cell type is necessary in order to determine if exosomes can be used for immunotherapy. Alveolar macrophages are immune cell that play a role in the defense against pathogens and clearing lung pathways. In order to confirm that exosomes could be isolated from J774 cells, a murine alveolar macrophage cell line, Western blotting was performed using CD63 antibodies as well as Nanoparticle Tracking Analysis. The molecule of interest packaged in the exosomes is the mRNA for Cystic Fibrosis Transmembrane Conductance Regulator or CFTR, an ABC transporter ion channel that is involved with cystic fibrosis. This channel plays an important role in the function of alveolar macrophages. Primer sets that span exons 9 through 13 of CFTR were used to confirm the presence of the CFTR mRNA inside the J774 cells as well as in exosomes. Further experiments are needed to determine if there are additional modulators for CFTR that are packaged with the mRNA as well as what changes in expression are seen when introduced into a knockout or mutated cell line.
Stanic, Purdue University.
Biology|Cellular biology|Molecular biology
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