title: Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days.
abstract: Autologous stem cell transplantation using unprocessed, G-CSF-mobilized whole blood (WB) is a simple, cost-reducing procedure and supports high-dose chemotherapy regimens not exceeding 72 h. Thereafter, clonogenic capacity rapidly decreases if routine anticoagulants are used for storage. In order to increase clinical applicability, we investigated the requirements for optimal preservation of unprocessed WB for 7 days. During storage at 22 degrees C in CPDA-1, a decrease in pH was noted, which was at least partially responsible for the low recovery of clonogenic cells. Subsequently, WB cells were stored in various cell culture media (RPMI 1640, alpha-MEM, X-VIVO15, CellGro SCGM and Leibovitz's L15 medium) containing either serum, serum-free substitutes or no additives. Leibovitz's L15 showed significantly better CFU-GM recoveries than the other media. Using a calcium-free modification of L15 medium (added 3:10 to WB), 94 +/- 24% of CD34(+) cells, 41 +/- 14% of BFU-E, 56 +/- 17% CFU-GM and 90 +/- 14% of LTC-IC were preserved during storage for 7 days at 22 degrees C. Storage at 4 degrees C was also feasible, but showed less optimal recoveries of 52 +/- 29% (CD34), 32 +/- 10% (BFU-E), 13 +/- 7% (CFU-GM) and 58 +/- 9% (LTC-IC). The expression of CD38, Thy-1, c-kit, AC133, L-selectin and CXCR4 on CD34-positive cells remained unchanged. In conclusion, a modified Leibovitz's L15 medium better meets the metabolic requirements of a high-density cell culture and allows safe storage of G-CSF mobilized WB for at least 7 days. The results encourage further exploration of WB transplants stored for 7 days for clinical use.
authors.affiliation: Department of Hematology, VU Medical Center, Amsterdam, The Netherlands.
authors.forename: A M
authors.initials: AM
authors.lastname: de Kreuk
authors.forename: A R
authors.initials: AR
authors.lastname: Jonkhoff
authors.forename: A
authors.initials: A
authors.lastname: Zevenbergen
authors.forename: E C
authors.initials: EC
authors.lastname: Hendriks
authors.forename: G J
authors.initials: GJ
authors.lastname: Schuurhuis
authors.forename: G J
authors.initials: GJ
authors.lastname: Ossenkoppele
authors.forename: A M
authors.initials: AM
authors.lastname: Dräger
authors.forename: J W
authors.initials: JW
authors.lastname: van Oostveen
authors.forename: P C
authors.initials: PC
authors.lastname: Huijgens
mesh_terms: D000225:Adenine; D015703:Antigens, CD / Q000097:blood; D018952:Antigens, CD34 / Q000097:blood; D001793:Blood Preservation / Q000379:methods*; D001943:Breast Neoplasms / Q000628:therapy; D018929:Cell Culture Techniques / Q000379:methods; D002951:Citrates; D003114:Colony-Forming Units Assay; D003470:Culture Media; D016895:Culture Media, Serum-Free; D005260:Female; D000069585:Filgrastim; D005947:Glucose; D016179:Granulocyte Colony-Stimulating Factor / Q000627:therapeutic use*; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D000078224:Lenograstim; D008228:Lymphoma, Non-Hodgkin / Q000628:therapy; D009369:Neoplasms / Q000628:therapy*; D009930:Organic Chemicals; D010710:Phosphates; D011994:Recombinant Proteins / Q000627:therapeutic use*; D014182:Transplantation, Autologous
chemical_list: D015703:Antigens, CD; D018952:Antigens, CD34; C034876:CPDA solutions; D002951:Citrates; D003470:Culture Media; D016895:Culture Media, Serum-Free; C095240:L15 culture medium; D009930:Organic Chemicals; D010710:Phosphates; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D000078224:Lenograstim; D005947:Glucose; D000225:Adenine; D000069585:Filgrastim