Human Mesenchymal Stem Cell-Derived Giant Plasma Membrane Vesicles(GPMV) As A Novel Candidate for Liver Regeneration

Abstract

Giant plasma membrane vesicles (GPMV) are micron-sized vesicles derived from the plasma membrane of mammalian cells under the influence of chemical vesiculants. In this work we have made an attempt to explore the regenerative potential of mesenchymal stem cell derived GPMV. For this purpose, in vitro cell culture based drug-induced liver disease model was adopted. We synthesized GPMV from adipose tissue derived mesenchymal stem cell using chemical vesiculants namely dithiothreitol (DTT) and paraformaldehyde (PFA). The synthesized GPMVs were found in the size range of 2-15µm. We confirm the presence of cytosol in the GPMV by calcein-AM staining. Fluorescence recovery after photo bleaching assay further showed that solute mobility inside the GPMV was almost 2 fold higher in comparison to that of inside the stem cell. Using a set of dyes, we prove that GPMVs contain proteins and RNA thus can work as packaged cellular cargo for the delivery of bioactive molecules but are devoid of DNA. The proteins and RNA are homogeneously distributed. To evaluate the regenerative potential of GPMVs, we created an acetaminophen (APAP) induced diseased liver model in vitro. The restoration of the functionality of the APAPtreated cells under the influence of GPMV was evaluated by checking the expression of albumin by immunocytochemistry. This study suggests that the mesenchymal stem cell derived GPMV can be used as a new stem cell derived therapeutics.