# US 2020/0399607 A1 · System and method for extraction and cryopreservation of bone marrow > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2020/0399607 A1 - Document ID: US20200399607A1 - Status: Published application - Kind: A1 - Application number: 17/013,400 - Filed: 2020-09-04 - Published: 2020-12-24 - Earliest claimed: 2019-04-15 - Assignees: Ossium Health, Inc. - Applicants: Ossium Health, Inc. - Inventors: Erik John WOODS; Brian H. JOHNSTONE; Dongsheng GU; Aubrey Marie SHERRY; Kelsey Gwen MUSALL; John R. WOODS; James HARDIN; Alan HOOKS - Web page: https://patentgrep.com/patent/US20200399607A1 ## CPC - C12N 5/0663: {Bone marrow mesenchymal stem cells (BM-MSC)} - A01N 1/125: Freeze protecting agents, e.g. cryoprotectants or osmolarity regulators - A61B 17/1611: {the two jaw elements being integral with respective elongate shaft elements} - A61K 35/28: Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells - C12M 45/02: {by mechanical forces; Stirring; Trituration; Comminuting (crushing, pulverizing, disintegrating in general B02C)} - C12M 45/05: {by centrifugation (centrifuges in general B04B)} - A61B 17/1606: {of forceps type, i.e. having two jaw elements moving relative to each other} - A61B 2017/00831: {Material properties} - A61B 2017/00969: {used for transplantation} - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) - A01N: PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF (preservation of food or foodstuff A23); BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES (preparations for medical, dental or toiletry purposes which kill or prevent the growth or proliferation of unwanted organisms A61K); PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS - A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D) - A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D) - C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS} ## 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. ## Claims ### Claim 1 A method for recovering vertebral bone adherent mesenchymal stromal/stem cells (vBA-MSC) from cadaver bone or cadaver bone fragments, the method comprising: (a) obtaining cadaver bone, cadaver bone fragments, or ground cadaver bone, optionally, preparing ground cadaver bone from the cadaver bone or cadaver bone fragments; (b) incubating the ground cadaver bone in a digestive solution comprising collagenase and/or neutral protease thereby obtaining a digested bone product; and (c) collecting the extracted bone marrow cells, thereby recovering the vBA-MSC. ### Claim 2 (canceled) ### Claim 3 (depends on 1) The method of claim 1, wherein the incubating is up to about 2.5 hours. ### Claim 4 (depends on 1) The method of claim 1, wherein the amount of neutral protease amount is about 20 U/ml. ### Claim 5 (canceled) ### Claim 6 (canceled) ### Claim 7 (depends on 1) The method of claim 1, wherein the collagenase is DE collagenase (Vitacyte), the collagenase comprises a recombinant collagenase isoform C1, and/or the collagenase comprises a recombinant collagenase isoform C2. ### Claim 8 (depends on 1) The method of claim 1, wherein the collagenase is recombinant Clostridium histolyticum collagenase. ### Claim 9 (depends on 1) The method of claim 1, wherein the neutral protease is Paneibacillus polymyxa neutral protease. ### Claim 10 (depends on 1) The method of claim 1, wherein the digestive solution comprises collagenase and neutral protease. ### Claim 11 (depends on 1) The method of claim 1, wherein the volume to weight ratio of the digestive solution to weight of the ground cadaver bone is about 5 to 1. ### Claim 12 (depends on 1) The method of claim 1, wherein ground cadaver bone is prepared from cadaver bone or cadaver bone fragments by grinding the cadaver bone or cadaver bone fragments with a bone grinder in the presence of a grinding medium. ### Claim 13 (depends on 12) The method of claim 12, wherein the grinding medium comprises two or more of Benzonase®, human serum albumin (HSA), heparin, and PLASMA-LYTE A. ### Claim 14 (depends on 13) The method of claim 13, wherein the ground cadaver bone is captured on a filter or sieve before the incubating in step (b) and filtrate that passes thru the filter or sieve comprises bone marrow cells. ### Claim 15 (depends on 14) The method of claim 14, further comprising collecting the filtrate comprising bone marrow cells. ### Claim 16 (depends on 14) The method of claim 14, wherein the captured ground bone is incubated in the digestive solution in step (b). ### Claim 17 (depends on 16) The method of claim 16, wherein the digestive solution is neutralized by providing a human platelet lysate. ### Claim 18 (depends on 16) The method of claim 16, further comprising steps of filtering the ground bone after incubating, washing the ground bone after filtering after incubating, and/or collecting filtrate after incubating, wherein the filtrate comprises vBA-MSCs. ### Claim 19 (depends on 17) The method of claim 17, wherein the filtrate comprising vBA-MSCs is combined with the filtrate comprising bone marrow cells or the filtrate comprising vBA-MSCs is not combined with the filtrate comprising bone marrow cells. ### Claim 20 (depends on 1) The method of claim 1, wherein the vBA-MSCs are phenotypically and functionally equivalent to MSCs obtained from central bone marrow. ### Claim 21 (depends on 1) The method of claim 1, wherein the cadaver bone or the cadaver bone fragments are derived from a vertebral body, an ileum, or a combination thereof. ### Claim 22 (depends on 1) The method of claim 1, wherein the cadaver bone or cadaver bone fragments are derived from a plurality of cadaver bones from the same donor. ### Claim 23 (depends on 1) The method of claim 1, wherein the method provides more than 2 trillion vBA-MSC cells per 100 g of cadaver bone or cadaver bone fragments.