The establishment of a bank of stored clinical bone marrow stromal cell products.
BACKGROUND
Bone marrow stromal cells (BMSCs) are being used to treat a variety of conditions. For many applications a supply of cryopreserved products that can be used for acute therapy is needed. The establishment of a bank of BMSC products from healthy third party donors is described.


METHODS
The recruitment of healthy subjects willing to donate marrow for BMSC production and the Good Manufacturing Practices (GMP) used for assessing potential donors, collecting marrow, culturing BMSCs and BMSC cryopreservation are described.


RESULTS
Seventeen subjects were enrolled in our marrow collection protocol for BMSC production. Six of the 17 subjects were found to be ineligible during the donor screening process and one became ill and their donation was cancelled. Approximately 12 ml of marrow was aspirated from one posterior iliac crest of 10 donors; one donor donated twice. The BMSCs were initially cultured in T-75 flasks and then expanded for three passages in multilayer cell factories. The final BMSC product was packaged into units of 100 × 106 viable cells, cryopreserved and stored in a vapor phase liquid nitrogen tank under continuous monitoring. BMSC products meeting all lot release criteria were obtained from 8 of the 11 marrow collections. The rate of growth of the primary cultures was similar for all products except those generated from the two oldest donors. One lot did not meet the criteria for final release; its CD34 antigen expression was greater than the cut off set at 5%. The mean number of BMSC units obtained from each donor was 17 and ranged from 3 to 40.


CONCLUSIONS
The production of large numbers of BMSCs from bone marrow aspirates of healthy donors is feasible, but is limited by the high number of donors that did not meet eligibility criteria and products that did not meet lot release criteria.
Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Building 10, Room 1C711, Bethesda, MD 20892-1184, USA.
Marianna
M
Sabatino
Jiaqiang
J
Ren
Virginia
V
David-Ocampo
Lee
L
England
Michael
M
McGann
Minh
M
Tran
Sergei A
SA
Kuznetsov
Hanh
H
Khuu
Arun
A
Balakumaran
Harvey G
HG
Klein
Pamela G
PG
Robey
David F
DF
Stroncek
D000328:Adult; D000368:Aged; D000375:Aging / Q000502:physiology; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology*; D002452:Cell Count; D002467:Cell Nucleus / Q000378:metabolism; D049109:Cell Proliferation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005260:Female; D006262:Health; D006801:Humans; D008297:Male; D008875:Middle Aged; D017154:Stromal Cells / Q000166:cytology; D013997:Time Factors; D014015:Tissue Banks; D014019:Tissue Donors; D014021:Tissue Preservation / Q000379:methods*; D055815:Young Adult
D018952:Antigens, CD34