System and methods for preparation of adipose-derived stem cells

A system for processing tissue in a closed pathway for maintaining sterility comprising:

• a first centrifuge tube having an essentially cylindrical body having a top, an inverse conical apex and an interior volume, the first centrifuge tube further comprising:
• at least two swabable valves, the at least two swabable valves comprising at least one extraction fitting;
• an extraction tube affixed to the at least one extraction fitting that extends a distance within the interior volume towards the apex;
• a screen disposed within the interior volume of the body;
• and
• a filter positioned below the screen in the inverse conical apex of the first centrifuge tube;
• and
• a second centrifuge tube comprising an essentially cylindrical body having a top, an inverse conical apex, and an interior volume, the second centrifuge tube further comprising at least one swabable valve, and an extraction tube releasably affixed to the at least one swabable valve that extends a distance within the interior volume towards the apex,
• wherein the second centrifuge tube is releasably connected via the at least one swabable valve to the at least one extraction fitting of the first centrifuge tube.

A multiple unit enabled system for processing tissue in a closed pathway for maintaining sterility comprising:

• a centrifuge tube having an essentially cylindrical body having a top, an inverse conical apex and an interior volume and further comprising:
• at least two swabable valves, the at least two swabable valves comprising at least one extraction fitting,
• an extraction tube affixed to the at least one extraction fitting that extends a distance within the interior volume towards the apex,
• a screen disposed within the interior volume of the body, and
• a filter being positioned below the screen in the inverse conical apex of the centrifuge tube;
• and,
• a container having at least one swabable valve,
• wherein the swabable valve can be releasably connected to the at least one extraction fitting of the centrifuge tube for a sterile transfer of material contained with the closed pathway.

A method for preparation of a fat graft, the method comprising the steps of:

• (a) performing a sterile transfer of lipoaspirate to a centrifuge tube comprising an upper cylindrical portion and a lower conical portion, a sterile tissue inlet fitting, at least one sterile processing fluid inlet fitting, a sterile suction fitting, and at least one sterile extraction port connected to a first extraction tube, the centrifuge tube further comprising a screen being positioned therein, and a filter positioned therein, the filter being positioned below the screen in the lower conical portion of the centrifuge tube;
• (b) washing the lipoaspirate in the centrifuge tube by adding one or more fluids via the at least one sterile processing fluid inlet fitting;
• (c) centrifuging the centrifuge tube containing the lipoaspirate at a low speed to separate contaminants and dense fat fraction, where the dense fat fraction remains on top of the screen and liquids are collected in the lower conical portion;
• (d) centrifuging the centrifuge tube containing the dense fat fraction at a higher speed to force at least a part of the dense fat fraction through pores of the screen by a centrifuge force in order to accumulate fat particles in a compartment between the screen and the filter, wherein a surface of the filter serves as a bottom for the fat particles, and wherein debris filter down through the filter to the lower conical portion;
• (e) collecting at least some of the fat particles by a syringe, and by connecting the syringe to at least one of the inlet with corresponding tubing leading to the compartment between the filter and the screen;
• and
• (f) administering the at least some of the fat particles to a patient for a therapeutic or cosmetic purpose.

A centrifuge tube comprising:

• a body comprising a generally cylindrical portion and a conical portion having a bottom;
• a screen disposed within the body, the screen including a plurality of pores, the screen configured to support at least a part of a biological substance;
• a filter positioned in the conical portion and closer to the bottom of the conical portion than the screen, the filter including a plurality of pores, the plurality of pores for the filter being smaller than the plurality of pores for the screen;
• at least one tissue inlet fitting configured to introduce the biological substance into the generally cylindrical portion;
• and
• at least one extraction port connected to tubing, an end of the tubing positioned in a section of the body between the screen and the filter;
• wherein the plurality of pores of the screen are sized such that upon centrifugation, viscous tissue of the biological substance is broken down into smaller particles;
• and
• wherein the plurality of pores of the filter are sized such that upon centrifugation, a liquid fraction from the smaller particles is moved downward below the filter so that micronized tissue is positioned between the screen and the filter and supported by the filter.