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-11447750-B2
Publication Date
2022-09-20
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 provides a method for recovering vertebral bone marrow mesenchymal stromal/stem cells (vBM-MSCs) and vertebral bone adherent mesenchymal stromal/stem cells (vBA-MSCs) from cadaver bone or cadaver bone fragments. Ground cadaver bone is contacted with a grinding medium under conditions sufficient to separate bone marrow comprising vBM-MSCs from the ground cadaver bone.
A filtrate that passes through a filter or sieve is collected to recover the bone marrow comprising vBM-MSCs. The byproduct captured ground cadaver bone is then incubated in a digestive solution comprising collagenase and neutral protease, and the liberated vBA-MSCs are collected.
The method includes a yield limitation that at least 2×10^8 vBA-MSC are obtained from 100 g of captured byproduct ground cadaver bone. The overall approach focuses on recovering both vBM-MSCs and vBA-MSCs using a two-stage separation and digestion sequence specifically defined for vertebral bone-derived material.
Claims Coverage
The provided claim material includes clm-00001. The main inventive features are concentrated on a defined order of separating vBM-MSC-containing marrow and capturing the byproduct, followed by enzymatic liberation of vBA-MSCs from the captured byproduct, with an explicit yield constraint for vBA-MSC recovery.
Sequential recovery of vBM-MSCs and vBA-MSCs from ground vertebral cadaver bone by filtration and collagenase/neutral protease digestion
A method recovering vertebral bone marrow mesenchymal stromal/stem cells (vBM-MSCs) and vertebral bone adherent mesenchymal stromal/stem cells (vBA-MSC) from cadaver bone or cadaver bone fragments by contacting ground cadaver bone with a grinding medium to separate vBM-MSC-containing marrow, capturing a byproduct ground cadaver bone on a filter or sieve and collecting a filtrate passing through the filter or sieve, then incubating the captured byproduct ground cadaver bone in a digestive solution comprising collagenase and neutral protease to liberate vBA-MSCs, and collecting the liberated vBA-MSCs, wherein steps (b) and (c) precede step (d).
Minimum vBA-MSC yield from captured byproduct ground cadaver bone
The sequential method further specifies that at least 2×10^8 vBA-MSC are obtained from 100 g of captured byproduct ground cadaver bone.
The inventive coverage centers on a sequential workflow where vBM-MSC-containing marrow is separated and recovered by grinding followed by filtration or sieving, and the remaining captured byproduct is subsequently digested with collagenase and neutral protease to liberate and recover vBA-MSCs, with an explicit minimum yield requirement for vBA-MSC recovery.
Stated Advantages
Provides recovery of both vertebral bone marrow mesenchymal stromal/stem cells (vBM-MSCs) and vertebral bone adherent mesenchymal stromal/stem cells (vBA-MSCs) from cadaver bone or cadaver bone fragments.
Achieves a defined vBA-MSC yield of at least 2×10^8 vBA-MSC from 100 g of captured byproduct ground cadaver bone.
Documented Applications
No documented applications found
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