System and method for extraction and cryopreservation of bone marrow
Inventors
Woods, Erik John • Johnstone, Brian H. • GU, Dongsheng • Sherry, Aubrey Marie • Musall, Kelsey Gwen • Woods, John R. • Hardin, James • Hooks, Alan
Assignees
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Publication Number
US-11104882-B2
Publication Date
2021-08-31
Expiration Date
Abstract
Methods are provided for extracting bone marrow cells from bone obtained from deceased donors, for preparing the bone marrow for cryopreservation and for obtaining desired cells from cryopreserved and fresh bone marrow.
Core Innovation
The invention relates to a method for high-yield recovery of stem cells from cadaver bone or cadaver bone fragments. Cadaver bone or cadaver bone fragments are processed to extract bone marrow cells, which are then combined with a reduced density solution mixture that forms a density gradient. The density gradient is provided by a hydrophilic copolymer of sucrose and epichlorohydrin together with an electrolyte medium.
The extracted bone marrow cells are centrifuged to separate out buffy coat cells comprising stem cells. The buffy coat cells are harvested to recover stem cells comprising hematopoietic stem cells and/or mesenchymal stem cells. The approach is configured around collecting a stem-cell-containing buffy coat fraction after density-gradient separation using the reduced density solution mixture.
The reduced density solution mixture provides a density gradient of 1.063-1.052 g/m, and the buffy coat fraction is further defined by positional or density constraints in the density gradient. Further recovery steps isolate a CD34+ cell population from the recovered stem cells.
Claims Coverage
The independent claim is clm-00001. It covers a density-gradient method using a reduced density sucrose-epichlorohydrin copolymer solution to recover a buffy coat fraction containing hematopoietic stem cells and/or mesenchymal stem cells, with dependent claim features including centrifugation constraints and CD34+ enrichment.
Reduced-density sucrose-epichlorohydrin density gradient in electrolyte medium
Obtaining a reduced density solution mixture of a hydrophilic copolymer of sucrose and epichlorohydrin at density 1.077 g/mL, mixed with an electrolyte medium, wherein the reduced density solution mixture provides a density gradient of 1.063-1.052 g/m.
Buffy coat separation by centrifugation of marrow cells combined with reduced density gradient
Combining the extracted bone marrow cells with the reduced density solution mixture; and centrifuging the extracted bone marrow cells, thereby separating out buffy coat cells comprising stem cells.
Harvesting buffy coat to recover hematopoietic and/or mesenchymal stem cells
Harvesting the buffy coat cells, thereby recovering stem cells comprising hematopoietic stem cells and/or mesenchymal stem cells.
Overall, claim coverage centers on forming the specified reduced-density sucrose-epichlorohydrin density gradient with an electrolyte medium, using centrifugation to separate a buffy coat containing stem cells, and harvesting that buffy coat to recover hematopoietic and/or mesenchymal stem cells.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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