title: A technique of bone marrow collection from vertebral bodies of cynomolgus macaques for transplant studies.
abstract: BACKGROUND
  Strategies to induce donor-specific allograft tolerance are best tested in preclinical models developed in nonhuman primates (NHPs). Most protocols prepare the recipient by infusing hematopoietic cells from the donor. We report here a procedure to isolate and characterize large numbers of bone marrow cells (BMCs) from cynomolgus monkeys (cynos) that can then successfully be transplanted into conditioned recipients.
  
  
  METHODS
  Vertebral columns of five cynos were excised en bloc and separated into individual vertebrae. The cancelous bone was extracted with a core puncher, fractionated, filtered, centrifuged, and resuspended in transplantation media before being analyzed by flow cytometry. In two instances, the collected BMCs were reinfused into allogeneic recipients preconditioned with a nonmyeloablative regimen. Chimerism was monitored using short-tandem repeat analysis.
  
  
  RESULTS
  The mean total BMCs yield was 25.5 x 10(9) (range of 4.00 x 10(9) to 59 x 10(9)) with mean cell viability of 93.4% (range: 90-96%). CD34+ cells and CD3+ cells averaged 0.34 and 3.91% of total BMCs, respectively. This resulted in absolute cell number yields of 1.02 x 10(8) and 1.15 x 10(9) for CD34+ and CD3+ cells, respectively. Graft-versus-host disease was absent in both bone marrow infused animals, and a maximum level of chimerism of 18% was detected at 3 weeks after BMCs infusion.
  
  
  CONCLUSIONS
  We present here the first detailed report of a procedure to retrieve and characterize large numbers of BMCs from vertebral bodies of cynos and demonstrate that cells collected with this technique have the capability of engrafting in allogenic recipients.
authors.affiliation: Transplantation Immunology Laboratory, Department of Cardiothoracic Surgery, Falk Cardiovascular Research Center, Stanford University School of Medicine, Stanford, CA 94305-5407, USA.
authors.forename: Mona G
authors.initials: MG
authors.lastname: Flores
authors.forename: Bari
authors.initials: B
authors.lastname: Holm
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Larson
authors.forename: Macy K
authors.initials: MK
authors.lastname: Lau
authors.forename: Ming-Sing
authors.initials: MS
authors.lastname: Si
authors.forename: Robert
authors.initials: R
authors.lastname: Lowsky
authors.forename: Geraldine
authors.initials: G
authors.lastname: Rousvoal
authors.forename: F Carl
authors.initials: FC
authors.lastname: Grumet
authors.forename: Samuel
authors.initials: S
authors.lastname: Strober
authors.forename: Richard
authors.initials: R
authors.lastname: Hoppe
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Reitz
authors.forename: Dominic C
authors.initials: DC
authors.lastname: Borie
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology* / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D006084:Graft Rejection / Q000188:drug therapy; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000276:immunology; D007108:Immune Tolerance; D007166:Immunosuppressive Agents / Q000494:pharmacology; D008252:Macaca fascicularis; D008297:Male; D013131:Spine / Q000166:cytology*; D018183:Transplantation Chimera; D014184:Transplantation, Homologous
chemical_list: D007166:Immunosuppressive Agents