| CPC C12M 45/09 (2013.01) [C12M 23/28 (2013.01); C12M 23/44 (2013.01); C12M 27/02 (2013.01); C12M 27/10 (2013.01); C12M 33/00 (2013.01); C12M 33/14 (2013.01)] | 20 Claims |
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1. A method for processing biological material including adipose tissue to dissociate the adipose tissue and release cellular components from the adipose tissue for preparation of stromal vascular fraction, the method comprising:
introducing an aqueous digestion medium into an internal fluid containment space of a portable, single-use processing container of a portable tissue processing unit to contact biological material including adipose tissue in the internal fluid containment space, wherein the digestion medium includes enzyme for enzymatic digestion of the adipose tissue and wherein the tissue processing unit comprises:
an axis of rotation extending through the processing container in a direction from a bottom of the processing container toward a top of the container, the tissue processing unit and the processing container being rotatable about the axis of rotation for rotational processing of contents in the internal fluid containment space;
at least one selectively sealable and un-sealable inlet port through the top of the processing container and in fluid communication with the internal fluid containment space;
an upright orientation and a reclined orientation, wherein in the reclined orientation the axis of rotation is at a reclined angle to horizontal relative to the upright orientation and wherein in the reclined orientation the axis of rotation is inclined relative to horizontal at an angle of inclination in a range of from 0° to 45°; and
at least one mixing blade disposed in the internal fluid containment space of the processing container;
during the introducing the digestion medium, the tissue processing unit is in the upright orientation and the digestion medium is introduced into the internal fluid containment space through an unsealed said inlet port;
after the introducing the digestion medium, enzymatically digesting the adipose tissue of the biological material in the internal fluid containment space in presence of the enzyme in the internal fluid containment space to dissociate the adipose tissue and release cells of stromal vascular fraction from the adipose tissue, comprising rotating the tissue processing unit and the processing container about the axis of rotation while the tissue processing unit is in the reclined orientation with the said inlet port sealed;
after the introducing the digestion medium and prior to the rotating, repositioning the tissue processing unit from the upright orientation to the reclined orientation with the biological material and the digestion medium in a process material mass in the internal fluid containment space in the reclined orientation, wherein in the reclined orientation the process material mass is in an amount having a quiescent volume corresponding to a quiescent partial fill volume of the internal fluid containment space with a quiescent headspace volume in the internal fluid containment space above the quiescent partial fill volume; and
the rotating the tissue processing unit and the processing container comprising:
rotating the processing container and the at least one mixing blade together as a unit and not relative to each other; and
during the rotating the processing container and the at least one mixing blade together as a unit and not relative to each other, moving the at least one mixing blade through periodically repeating mixing cycles in the internal fluid containment space to contact and mix the process fluid mass, wherein each said mixing cycle comprises moving at least a portion of the at least one mixing blade sequentially through the quiescent partial fill volume and the quiescent headspace volume.
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