Bone marrow adipose portion isolation device and methods
Inventors
Centeno, Christopher • Dregalla, Ryan • Lyons, Nicolette • Reischling, Patrick
Assignees
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Publication Number
US-10537596-B2
Publication Date
2020-01-21
Expiration Date
Abstract
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.
Core Innovation
The invention relates to systems, devices, and methods for processing bone marrow aspirate by fractionating the aspirate within a first chamber into layers including an adipose layer supernatant. Instead of discarding the adipose layer supernatant, the processed adipose layer supernatant is collected for further processing, with device configurations that include first and second chambers in fluid communication for transferring and handling the adipose layer supernatant.
The collected adipose layer supernatant is processed in a mechanical emulsification system in communication with the second chamber. The mechanical emulsification system includes a first emulsification chamber and a second emulsification chamber arranged to emulsify upon passage of the adipose layer supernatant, including configurations using an aperture or an emulsification screen. The mechanical emulsification step dissociates mesenchymal stem cells (MSCs) from an adipose collagen matrix.
After emulsification, the invention describes processing the emulsified adipose layer supernatant to collect mesenchymal stem cells. The document further includes optional addition of secondary substances or digestion agents such as collagenase and lecithin, and/or application of sonic energy or vibration. The disclosed workflows and device variants are reported to yield markedly higher MSC yields and improved culture performance compared with buffy coat processing, including a stated ~700% increase.
Claims Coverage
The independent claim recites a method centered on fractionating bone marrow aspirate to obtain an adipose layer supernatant in a first chamber, collecting that adipose layer supernatant in a second chamber, and emulsifying the adipose layer supernatant using a mechanical emulsification system. Dependent claim coverage refines the emulsification system structure and adds options for downstream MSC collection processing.
Two-chamber fractionation with adipose layer supernatant collection
A device is provided with a first chamber and a second chamber in fluid communication, wherein bone marrow aspirate is fractionated within the first chamber into layers including an adipose layer supernatant, the adipose layer supernatant is collected from the processed bone marrow aspirate in the second chamber, and the adipose layer supernatant is directed to mechanical emulsification.
Mechanical emulsification of adipose layer supernatant
A mechanical emulsification system in fluid communication with the second chamber emulsifies the collected adipose layer supernatant.
Aperture-based emulsification between chambers
The mechanical emulsification system comprises a first emulsification chamber and a second emulsification chamber in fluid communication through an aperture sized to provide emulsification upon passage of the adipose layer supernatant between the first and second emulsification chambers.
Emulsification screen-based emulsification between chambers
The mechanical emulsification system comprises a first emulsification chamber and a second emulsification chamber in fluid communication through an emulsification screen, wherein emulsification occurs upon passage of the adipose layer supernatant between the first and second emulsification chambers.
Downstream processing to collect mesenchymal stem cells from adipose layer supernatant
The method further comprises processing the adipose layer supernatant to collect mesenchymal stem cells.
Adding a digestion agent to the collected adipose layer supernatant
The method further comprises adding a digestion agent to the collected adipose layer supernatant.
Fractionation and separation structures within the first chamber
The first chamber comprises one or more of: a cap with a fluid access port, a plunger providing for expulsion of a selected portion, a disk shaped volume enabling collection of an adipose layer supernatant fraction at a central region upon rotation, a restricted-diameter portion for locating the adipose fraction, a floating density-selected disk floating between a serum layer and adipose layer supernatant fraction, a porous lipophilic membrane for separating the adipose layer supernatant fraction, and one or more ports in fluid communication with an adipose layer supernatant fraction.
Overall, the claim set is directed to collecting an adipose layer supernatant from fractionated bone marrow aspirate and emulsifying it with a mechanical emulsification system, with dependent claims narrowing the emulsification mechanism using an aperture or emulsification screen and adding downstream processing elements aimed at collecting mesenchymal stem cells, including optional digestion-agent addition and specific first-chamber fractionation/separation structures.
Stated Advantages
Markedly higher MSC yields, including a stated ~700% increase versus prior estimates.
Improved culture performance versus buffy coat processing.
Documented Applications
Processing bone marrow aspirate to collect mesenchymal stem cells (MSCs) for downstream culture performance comparisons, including comparisons to buffy coat processing.
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