Intra-operative preparation of autologous bone marrow-derived CD34-enriched cellular products for cardiac therapy.
OBJECTIVE
With the advent of regenerative therapy, there is renewed interest in the use of bone marrow as a source of adult stem and progenitor cells, including cell subsets prepared by immunomagnetic selection. Cell selection must be rapid, efficient and performed according to current good manufacturing practices. In this report we present a methodology for intra-operative preparation of CD34(+) selected autologous bone marrow for autologous use in patients receiving coronary artery bypass grafts or left ventricular assist devices.


RESULTS
We developed a rapid erythrocyte depletion method using hydroxyethyl starch and low-speed centrifugation to prepare large-scale (mean 359 mL) bone marrow aspirates for separation on a Baxter Isolex 300i immunomagnetic cell separation device. CD34 recovery after erythrocyte depletion was 68.3 ± 20.2%, with an average depletion of 91.2 ± 2.8% and an average CD34 content of 0.58 ± 0.27%. After separation, CD34 purity was 64.1 ± 17.2%, with 44.3 ± 26.1% recovery and an average dose of 5.0 ± 2.7 × 10(6) CD34(+) cells/product. In uncomplicated cases CD34-enriched cellular products could be accessioned, prepared, tested for release and administered within 6 h. Further analysis of CD34(+) bone marrow cells revealed a significant proportion of CD45(-) CD34(+) cells.


CONCLUSIONS
Intra-operative immunomagnetic separation of CD34-enriched bone marrow is feasible using rapid low-speed Hetastarch sedimentation for erythrocyte depletion. The resulting CD34-enriched product contains CD45(-) cells that may represent non-hematopoietic or very early hematopoietic stem cells that participate in tissue regeneration.
Department of Medicine, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA. donnenbergad@upmc.edu
Albert D
AD
Donnenberg
Vera S
VS
Donnenberg
Deborah L
DL
Griffin
Linda R
LR
Moore
Ferda
F
Tekinturhan
Robert L
RL
Kormos
D000368:Aged; D018952:Antigens, CD34 / Q000378:metabolism*; D001854:Bone Marrow Cells / Q000166:cytology* / Q000378:metabolism*; D001026:Coronary Artery Bypass / Q000379:methods; D005260:Female; D005434:Flow Cytometry; D006321:Heart; D006352:Heart Ventricles / Q000601:surgery; D006801:Humans; D007432:Intraoperative Period; D008297:Male; D008875:Middle Aged; D014182:Transplantation, Autologous / Q000379:methods*
D018952:Antigens, CD34