title: Collection and processing of umbilical cord blood for cryopreservation.
abstract: Umbilical cord blood (UCB) is being increasingly used as an alternative source of hematopoietic stem cells for allogeneic bone marrow transplantation. UCB transplantation has been successfully used to treat a variety of genetic, hematological, and oncological disorders in children and adults. The objectives of this study was to establish a closed-system technique for UCB collection and buffy coat separation by Optipress I device. Thirty-four UCB were collected by triple-bag system from pregnant mothers whose fetuses were not affected by thalassemic diseases after prenatal diagnosis. The mean volumn of UCB collection were 120 +/- 5 ml (range 65-180 ml). Total WBC, CD34+ cells, the progenitor cell erythroid burst-forming unit (BFU-E) and granulocyte-macrophage colony-forming unit (CFU-GM) in the UCB units were (9.36 +/- 0.84) x 10(8), (3.61 +/- 0.52) x 10(6), (9.12 +/- 1.60) x 10(5), and (5.32 +/- 1.23) x 10(5), respectively. Good correlation between the nucleated cell and net cord blood volume could be demonstrated (p < 0.0001). The correlation between CD34+ cells and the following parameters: nucleated cell, BFU-E or CFU-GM were also demonstrated (p = 0.001, 0.0105 or 0.0001, respectively). Buffy coat was subsequently separated from 18 UCB units by Optipress I device. 70 +/- 3 ml of buffy coat were collected and cryoprocessing was done by automatic controlled-rate freezer. Good recovery of total WBC, CD34+ cells, progenitor cells BFU-E and CFU-GM after buffy coat separation were observed 89 per cent, 95 per cent, 109 per cent, and 102 per cent respectively. There was no aerobic bacterial or fungal contamination in the separated blood products. By using this technique, the UCB units were easily collected, rapidly separated within one hour, and high recovery of the hematopoietic progenitor cells could be obtained.
authors.affiliation: Division of Hematology, Department of Medicine, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700, Thailand.
authors.forename: Yaowalak
authors.initials: Y
authors.lastname: U-pratya
authors.forename: Sirikwan
authors.initials: S
authors.lastname: Boonmoh
authors.forename: Orathai
authors.initials: O
authors.lastname: Promsuwicha
authors.forename: Chanya
authors.initials: C
authors.lastname: Theerapitayanon
authors.forename: Laksami
authors.initials: L
authors.lastname: Kalanchai
authors.forename: Viruch
authors.initials: V
authors.lastname: Chanjerboon
authors.forename: Korakot
authors.initials: K
authors.lastname: Sirimai
authors.forename: Sanan
authors.initials: S
authors.lastname: Visuthisakchai
authors.forename: Sasitorn
authors.initials: S
authors.lastname: Bejrachandra
authors.forename: Surapol
authors.initials: S
authors.lastname: Issaragrisil
mesh_terms: D018952:Antigens, CD34 / Q000032:analysis; D001793:Blood Preservation / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D015331:Cohort Studies; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D005312:Fetal Blood / Q000166:cytology* / Q000637:transplantation; D006412:Hematopoietic Stem Cells; D006801:Humans; D011247:Pregnancy; D012680:Sensitivity and Specificity; D013048:Specimen Handling; D013785:Thailand
chemical_list: D018952:Antigens, CD34