The Alterations of the Activities of Coagulation Inhibitors and Fibrinolytic Factors in Stored Cord Blood Could Affect the Yield of Progenitor Cells During Processing

Lee, Young-Ho; Han, Jin‐Yeong; Choi, Ahn-Hong; Lee, Hyung-Sik; Hur, Won-Joo; Seo, Suyoung; Han, Hun; Kim, Jae‐Seok; Kim, Hyojin

doi:10.1089/152581699319821

Cited by 7 publications

(7 citation statements)

References 7 publications

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“…Initially, the flasks were slowly agitated, but this led to significant clumping of the white cells at 4°C, so the agitation was omitted. 20 No significant differences in recovery of clonogenic cells were found between agitation and no agitation at 22°C (data not shown). Samples of 100 ml were stored at 22°C in Lifecell PL732 Culture Bags (Nexell Therapeutics Inc, Irvine, CA, USA).…”

Section: Storage Conditionsmentioning

confidence: 87%

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Kreuk

Jonkhoff

Zevenbergen

et al. 2001

Bone Marrow Transplant

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Summary: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؇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, ␣-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؇C. Storage at 4؇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. Bone Marrow Transplantation (2001) 28, 145-155.

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Section: Storage Conditionsmentioning

confidence: 87%

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Kreuk

Jonkhoff

Zevenbergen

et al. 2001

Bone Marrow Transplant

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“…In a previous study [12], we observed small clumps in the CB following red cell depletion, particularly in the samples stored longer than 24 hr. Microscopic findings of small clumps revealed aggregates of nucleated cells consisted of progenitor cells admixed with fibrin strands.…”

Section: Introductionmentioning

confidence: 57%

“…This phenomenon was related to the alterations of the activities of coagulation inhibitors and fibrinolytic factors in elapsed CB. In particular, the plasma activity of ␣ 2 antiplasmin, which is the most important fast-reacting plasmin inhibitor, decreased significantly in CB stored at ambient temperature for 48 hr [12].…”

Section: Discussionmentioning

confidence: 96%

Urokinase could increase the yield of progenitor cells during red cell depletion in elapsed and anticoagulated cord blood

et al. 2002

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We assessed whether the urokinase could increase the yield of progenitor cells during processing in elapsed, anticoagulated cord blood (CB) after collection, and we also determined the optimal dose of urokinase. The total nucleated cell (TNC) counts after red cell depletion in 48-hr-elapsed CB were significantly higher in samples treated with 10,000 and 50,000 IU of urokinase/mL than in untreated samples or treated with 5,000 IU of urokinase/mL. The CD34 + cell counts were significantly higher in samples treated with 10,000 IU of urokinase/mL than in untreated samples and in samples treated with 5,000 or 50,000 IU of urokinase/mL. In 6-, 12-, and 24-hr-elapsed CB, however, there were no significant differences of TNC, CD34 + cells, or CFU-GM counts between untreated samples and samples treated with 10,000 IU of urokinase/mL. These findings suggest that the addition of 10,000 IU of urokinase/mL before red cell depletion in 48-hr-elapsed, anticoagulated CB could increase the yield of progenitor cells. However, there are no advantages in using urokinase for processing CB prior to 24 hr after collection. Am. J. Hematol. 71:336-339, 2002.

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“…Most UCB banks collect UCB between Sunday and Friday to allow processing of the unit within 24 h, and a prolongation of the working period of the laboratory would significantly increase personnel costs [21]. Storage of UCB longer than 24 h would decrease the yield of nucleated cells and CD34+ cells [22]. The consent procedure at our institution usually takes place during the routine outpatient pregnancy visits around 4 weeks before the estimated date of delivery or when the patient in labor is pain-free due to an epidural anesthesia in the 1st stage of labor.…”

Section: Discussionmentioning

confidence: 99%

Problems in Umbilical Cord Blood Collection

Troeger¹,

Meyer-Monard

Thewis

et al. 2007

Transfus Med Hemother

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The goal of the Cord Blood Bank Basel is to provide umbilical cord blood (UCB) for allogeneic stem cell transplantations from unrelated donors. Our registry contains 1,044 units, 12 have been released for transplantation (1%). In the past collection mainly took place at the Women’s Hospital Basel, where only 10% of deliveries resulted in HLA-typed and frozen transplants. The aim of our study was to evaluate the high deferral rate. Materials and Methods: During a 4-week period we assessed every delivery at our hospital by a questionnaire on patient data, consent procedure, inclusion or exclusion criteria and reasons for performing or not performing UCB collection. Results: 164 women delivered at our institution (135 vaginal deliveries, 29 cesarean sections). Among these, 131 were eligible for autologous UCB donation, whereas 33 women had to be excluded due to their medical history (26), triplet delivery (1), post partum hemorrhage (1), congenital malformation (1) or chorioamnionitis (4). Of the eligible women, 17 refused donating UCB, and 29 women underwent a cesarean section. 11 women were admitted under painful contractions. 18 had to be rejected due to lack of laboratory facilities at the weekend. Finally, 43 UCB samples had been taken, 2 for private banking purposes (4.6%), 21 (48.8%) for the public bank, and 20 samples (46.5%) which had to be discarded due to a low cell count. Conclusions: Donor selection is efficiently performed before the collection process. The rate of donor deferral and UCB sample discard is high and comparable to other UCB banks. The yield of UCB transplants could be increased if UCB collection would be additionally performed during cesarean section and with increased laboratory facilities during the weekend.

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The Alterations of the Activities of Coagulation Inhibitors and Fibrinolytic Factors in Stored Cord Blood Could Affect the Yield of Progenitor Cells During Processing

Cited by 7 publications

References 7 publications

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Storage of unprocessed G-CSF-mobilized whole blood in a modified Leibovitz's L15 medium preserves clonogenic capacity for at least 7 days

Urokinase could increase the yield of progenitor cells during red cell depletion in elapsed and anticoagulated cord blood

Problems in Umbilical Cord Blood Collection

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