Mitochondrial Collection and Concentration, and Uses Thereof

摘要

Preparations and methods of producing a concentrated mitochondrial preparation using inverted phase centrifugation are disclosed. The methods include centrifuging a closed-bottom tube containing a heavy phase including a mitochondrial suspension in the portion of the tube proximal to the axis of rotation and a light phase in the portion of the tube distal to the axis of rotation. The centrifugation causes the suspension to form a concentrated mitochondrial layer at the interface between the heavy layer and the light layer, and the fluid at this interface is collected to produce the concentrated mitochondrial preparation. The concentrated mitochondrial preparation can be used for various applications, including in vitro fertilization applications.

主权项

1. A method for producing a concentrated mitochondrial preparation comprising the steps of:
(a) centrifuging a closed-bottom tube containing:
(i) a heavy phase comprising a mitochondrial suspension in the portion of the tube proximal to the axis of rotation;(ii) a light phase in the portion of the tube distal to the axis of rotation; and(iii) a phase interface between the heavy phase and the light phase; wherein the closed-bottom tube is centrifuged at a speed and for a time sufficient to cause mitochondria in the mitochondrial suspension to form a concentrated mitochondrial layer in the heavy phase at the interface between the light phase and the heavy phase, and without causing inversion of the light and heavy phases; and (b) collecting a volume of fluid comprising at least a portion of the concentrated mitochondrial layer to produce the concentrated mitochondrial preparation.