Bone marrow adipose portion isolation device and methods
The embodiments disclosed herein generally relate to systems, devices and methods for the fractionation, isolation, extraction and processing of the adipose supernatant layer of a bone marrow aspirate. In particular, the various embodiments relate to systems devices and methods of obtaining, utilizing and processing the adipose supernatant layer of a bone marrow aspirate as a source of mesenchymal stem cells.
1. A method of processing bone marrow aspirate comprising:
providing a device for processing bone marrow aspirate comprising:
a first chamber;
a second chamber in fluid communication with the first chamber; and
a mechanical emulsification system in fluid communication with the second chamber;
fractionating bone marrow aspirate within the first chamber of the device into layers including an adipose layer supernatant;
collecting the adipose layer supernatant from the processed bone marrow aspirate in the second chamber; and
emulsifying the adipose layer supernatant in the mechanical emulsification system.
2. The method of claim 1 further comprising processing the adipose layer supernatant to collect mesenchymal stem cells.
3. The method of claim 1 further comprising adding a secondary substance to the bone marrow aspirate.
4. The method of claim 3 wherein the secondary substance comprises at least one of a biologically inert fluid, CaCl 2 , thrombin, a clotting agent or a polymerization agent.
5. The method of claim 1 further comprising adding a digestion agent to the collected adipose layer supernatant.
6. The method of claims 5 wherein the digestion agent comprises at least one of collagenase or lecithin.
7. The method of claim 1 further comprising applying at least one of sonic energy or vibration to the adipose layer supernatant.
8. The method of claim 1 wherein the second chamber comprises at least one of a syringe, pipette or tube in fluid communication with the adipose layer supernatant.
9. The method of claim 1 wherein the first chamber comprises one or more of:
a cap comprising a fluid access port providing for the second chamber to be placed into fluid communication with the adipose layer supernatant;
a plunger providing for the expulsion of a selected portion of the fractionated bone marrow aspirate from the first chamber;
a disk shaped volume which provides for the collection of an adipose layer supernatant fraction at a central region of the disk shaped volume upon the rotation of the first chamber around a central axis;
a portion of restricted diameter positioned to correspond with the location of an adipose layer supernatant fraction upon fractionation of bone marrow aspirate placed within the first chamber;
a floating disk having a density selected to cause the disk to float substantially between a serum layer and an adipose layer supernatant fraction upon fractionation of the bone marrow aspirate;
a porous lipophilic membrane providing for the separation of an adipose layer supernatant fraction upon fractionation of the bone marrow aspirate; and
one or more ports in fluid communication with an adipose layer supernatant fraction upon fractionation of the bone marrow aspirate.
10. The method of claim 1 wherein the mechanical emulsification system comprises a first emulsification chamber and a second emulsification chamber in fluid communication with each other through an aperture sized to provide emulsification upon passage of the adipose layer supernatant between the first and second emulsification chambers.
11. The method of claim 1 wherein the mechanical emulsification system comprises a first emulsification chamber and a second emulsification chamber in fluid communication with each other through an emulsification screen providing for the emulsification of the adipose layer supernatant upon passage of adipose layer supernatant between the first and second emulsification chambers.
12. The method of claim 1 wherein the mechanical emulsification system comprises an emulsification screen movable with respect to the adipose layer and providing for the emulsification of the adipose layer supernatant.