Bone Marrow Transplantation, (1998) 22 , 193–196  1998 Stockton Press All rights reserved 0268–3369/98 $12.00 http:/ / www.stockton-press.co.uk/ bmt Case report Transplantation of unrelated cord blood cells S Weinreb 1 , JC Delgado 2 , OP Clavijo 2 , EJ Yunis 2 , L Bayer-Zwirello 3 , L Polanski 3 , L Deluhery 1 , G Cohn 3 , JT Yao 4 , TC Stec 4 , D Higby 1 and C Andrzejewski 4 Departments of 1 Medicine, 3 OB/GYN and 4 Pathology, Baystate Medical Center, Springfield, MA; and 2 Department of Cancer Immunology and AIDS, Dana-Farber Cancer Institute, Boston, MA, USA Summary: A 43-year-old woman with Philadelphia chromosome (Ph) positive chronic myelogenous leukemia in acute phase received high-dose chemotherapy followed by transfusion of 12 randomly selected units of umbilical cord blood. HLA analysis showed cells of one donor from day + 10 to day + 43 post-transfusion. This unit was HLA class II identical with that of the patient. Keywords: placental blood; unmatched; multiple donors; CML There is considerable interest in umbilical cord blood (UCB) transplants (for reviews see Refs 1–4). Most trans- plants have been from HLA-matched relatives. However, there is increasing use of HLA-matched unrelated donors (reviewed in Ref. 3). In 1972, Ende et al 5 transfused UCB from eight non-HLA unrelated donors to a 16-year-old with ALL previously treated with conventional chemotherapy. Success was claimed based on showing RBCs from one donor for 8 months. Shen et al 6 described giving multiple units of HLA-unmatched UCB to four patients with solid tumors. A patient with CML in accelerated phase was trans- planted with HLA-partially matched UCB from one donor by Laporte et al 7 who reported successful transplantation of the cord blood stem cells into a 26-year-old women. We report a patient who received 12 units of HLA-untyped UCB for CML in acute phase after high-dose chemo- therapy. Case report The patient, a 43-year-old female, was diagnosed with Ph chromosome-positive CML in 1991. She was treated with hydroxyurea and interferon until 1993 when accelerated phase developed. She then received total body irradiation (TBI) and cyclophosphamide followed by an autotrans- plant. In April 1996 accelerated phase recurred. She was Correspondence: Dr C Andrzejewski, Transfusion Medicine Service, Department of Pathology, Baystate Medical Center, 759 Chestnut Street, Springfield, MA 01199, USA Received 7 August 1996; accepted 10 March 1998 treated with cytarabine to which she responded briefly. Based on a previously approved Institutional Review Board (IRB) transplantation protocol involving unrelated UCB and after informed consent, the patient received busulfan (12 mg/kg) and cyclophosphamide (120 mg/kg) followed by infusion of 3.18 × 10 9 mononuclear cells obtained from UCB from 12 unrelated donors. On day + 1 we began GVHD prophylaxis consisting of cyclosporine and solumedrol. Early signs of engraftment were observed on day + 10 with islands of trilineage hema- topoiesis noted in a bone marrow biopsy specimen. There were no clinical features of GVHD. Increased bilirubin developed between days + 14 and + 17 with a peak value at 7.6 mg/dl. The patient was discharged in a stable condition on day + 21 with a WBC count of 2.6 × 10 9 /l. Fetal hemo- globinized RBC and RhD-positive RBC were detected in the blood between day + 17 and day + 21 (patient’s blood type O RhD-negative). Southern blot analysis of DNA from blood cells on day + 16 showed no bcr/abl rearrangement (previously detected). Growth factor therapy was stopped on day + 31 when the WBC count was 10 × 10 9 /l. The WBC continued to rise and was 8.5 × 10 9 /l on day + 53 with reappearance of Ph chromosome-positive cells. Hydroxy- urea and interferon were begun on day + 71 and she died on day + 90 of leukemia. Umbilical cord blood collecting and processing Units of UCB were collected after fetal delivery during the third stage of labor, with the placenta in situ by cannulation of the umbilical vein using a 16-gauge needle attached to blood transfer pack. The collected cord bloods were pro- cessed using a modification of the technique described by Rubinstein et al. 8 Briefly, this involved the transfer of the blood to multiple sterile 50 ml conical tubes (Baxter Healthcare, Scientific Products Division, McGaw Park, IL, USA) using sterile technique in a laminar flow hood. After centrifugation (1500 g for 10 min at room temperature), the plasma supernatant was separated, and an aliquot of plasma was sent for disease marker testing. After addition of enough normal saline to bring the volume of the packed red blood cell/buffy coat preparation to 50 ml, the cells were again centrifuged. After centrifugation, the ‘wash’ supernatant was discarded and the interface ‘buffy coat’ preparation was transferred to a new appropriately marked tube. Immediately 8 ml (final concentration 1% hetastarch)