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An Enzymatic Method to Rescue Mesenchymal Stem Cells from Clotted Bone Marrow Samples
Published on: April 12, 2015
Preparation of Plasma Membrane Vesicles from Bone Marrow Mesenchymal Stem Cells for Potential Cytoplasm Replacement
Li-Qun Xu1, Mei-Jia Lin1, Yun-Pan Li1
1Multidisciplinary Research Center, Shantou University.
Abstract:
We have previously reported on the generation of plasma membrane vesicles (PMVs) through the mechanical extrusion of mammalian cells. The fusion of PMVs with mitochondrial deficient Rho0 cells restored mitotic activity under normal culture conditions. Atherosclerosis, type 2 diabetes, Alzheimer's disease, and cancer are age-related diseases that have been reported to be associated with multiple mechanical and functional defects in the cytosol and organelles of a variety of cell types. Bone marrow mesenchymal stem cells (BMSCs) represent a unique cell population from the bone marrow that possess self-renewal capabilities while maintaining their multipotency. The supplementation of senescence cells with young cytoplasm from autologous BMSCs via the fusion of PMVs provides a promising approach to ameliorate or even reverse age-associated phenotypes. This protocol describes how to prepare PMVs from BMSCs via extrusion through a polycarbonate membrane with 3 µm pores, determine the existence of mitochondria and examine the maintenance of membrane potential within PMVs using a confocal microscope, concentrate PMVs by centrifugation, and carry out the in vivo injection of PMVs into the gastrocnemius muscle of mice.
Insights
Plasma membrane vesicles (PMVs) from bone marrow mesenchymal stem cells (BMSCs) can rejuvenate aged cells. Injecting these PMVs into mice muscle shows potential for reversing age-associated cellular defects.
Area of Science:
- Cell Biology
- Stem Cell Biology
- Gerontology
Background:
- Age-related diseases involve cellular defects in cytosol and organelles.
- Bone marrow mesenchymal stem cells (BMSCs) possess self-renewal and multipotency.
- Plasma membrane vesicles (PMVs) can be generated from mammalian cells.
Purpose of the Study:
- To describe a protocol for preparing and utilizing PMVs from BMSCs.
- To investigate the potential of BMSC-derived PMVs in ameliorating age-associated phenotypes.
- To assess the feasibility of in vivo PMV application.
Main Methods:
- Generating PMVs from BMSCs via extrusion through a 3 µm polycarbonate membrane.
- Confirming mitochondrial presence and membrane potential in PMVs using confocal microscopy.
- Concentrating PMVs by centrifugation and injecting them into mouse gastrocnemius muscle.
Main Results:
- Established a protocol for BMSC-derived PMV preparation.
- Demonstrated the presence of mitochondria and membrane potential within PMVs.
- Successfully performed in vivo injection of PMVs into mouse muscle tissue.
Conclusions:
- BMSC-derived PMVs offer a promising approach for cellular rejuvenation.
- This method could potentially reverse age-associated cellular phenotypes.
- The protocol provides a foundation for further research into PMV-based therapies.
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