Generating Plasma Membrane Vesicles from Bone Marrow Stem Cells

Abstract

Source: Xu, L. et al., Preparation of Plasma Membrane Vesicles from Bone Marrow Mesenchymal Stem Cells for Potential Cytoplasm Replacement Therapy. J. Vis. Exp. (2017)

This video demonstrates a method to generate plasma membrane vesicles from bone marrow stem cells by passing them rapidly through a membrane filter. These PMVs show potential for cytoplasm replacement therapy.

Protocol

1. Assembly of the Apparatus

1. To ensure sterility, turn on the ultraviolet (UV) light of a tissue culture hood for 30 min before use.
2. Unscrew a disposable 25 mm filter unit and submerge the cap and bottom of the unit into a 200 mL glass cup filled with 75% ethanol for 30 min. The unit is made of medical-grade polypropylene.
3. Pick up the cap and bottom of the unit using forceps, shake off the remaining ethanol by hand, and allow the parts to air dry for 10 min in the hood.
4. Wet a 19 mm polycarbonate membrane in phosphate-buffered saline (PBS) and place it carefully on top of the supporting matrix of the bottom of ....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
IsoporeTM membranes	Millipore	TSTP04700	3 mm pore
Disposable filter unit	Xinya, Shanghai, China	25 mm	Medical grade polypropylene
Insulin syringe	BD	328446	1 ml