Background

1. Results of the Cord Blood Transplantation Study (COBLT) Unrelated Donor Banking Program from Donor Screening to Characterization of Banked Units Joanne Kurtzberg, MD 1, Elizabeth L Wagner, MPH 2, John K Fraser, PhD 3, Mitch S Cairo, MD 4, LeeAnn Jensen PhD 5, Geoff Cohen MS 2, Shelly L Carter, ScD 2, Nancy A Kernan, MD 6 and *** 7. 1 Duke University Medical Center, Durham, North Carolina, 27705 ; 2 The EMMES Corporation, Rockville, Maryland, 20850 ; 3 University of California, Los Angeles, California, 90095 ; 4 Georgetown University Medical Center, Washington DC, 20007 ; 5 National Heart, Lung and Blood Institute, Bethesda, Maryland, 20817 ; 6 Memorial Sloan-Kettering Cancer Center, New York, New York, 10021 and 7 ***sponsored by the National Heart, Lung and Blood Institute, Bethesda, Maryland, 20817

2. Background • COBLT: The Cord Blood Transplantation Study – sponsored by the NHLBI • Banking study • Transplant Study • Initiated in 1996 and funded: • 1 Coordinating Center (EMMES) • 3 Cord Blood Banks • Choc/GT Mitch Cairo • Duke Joanne Kurtzberg • UCLA John Fraser • 7 Transplant Centers • 21 additional centers added over tenure of the study

3. Banking Study: Plan of Attack • Steering Committee: • Chair, Nancy Kernan, MD • Bank Directors • Coordinating & StatisticalCenter - EMMES • Representatives from NHLBI • Transplanters, OB, Ethicist, Lab Techs • Created and validated SOPs for program • Monthly meeting x 18 months

4. Standard Operating Procedures • SOPs • Donor screening and recruitment • Donor education and consent • Maternal donor medical history and testing • Collection and transportation • Processing and cryopreservation • Maternal Sample and Cord Blood Testing • Quarantine and release • Long term storage • Search Registry

5. SOPs - Continued • Standardization and validation of laboratory procedures • HLA by molecular techniques • Cell counts, viability, CFUs, CD34 and subsets, CD3,4,8, CD19, CD16/56 • Creation of internet based banking data system and search registry • Interlaboratory/bank QA/QC program with sample exchanges for testing, HLA typing, banking and thawing

6. Study Eligibility • Collections ex utero • UCBs >40 ml or 6x10e8 TNC • Initiation of processing within 48h of collection • Recovery of >60% of TNC post processing • +CFU growth • Negative maternal ID testing and maternal and family risk history • Negative homozygous hemoglobinopathy • All banked units passed a study algorithm defined by the steering committee

7. Timeline • Recruitment initiated in October, 1997 • Collections initiated in December, 1997 • SOPs published 1998 • Fraser et al, J Hematotherapy 7:521-561. • <www.emmes.com> • Collections completed June, 2001.

8. Total Total consenting collections Total Total Total and not recorded on 70% 85% 98% 83% Screened approached consented excluded monthly 34799 24240 20710 before recruitment delivery forms 20278 16764 15% 17% 30% 2% Total refusals 3530 Total not approached: 10559 Total collection failures at delivery: 3514 Reasons for not approaching Total excluded prior to delivery: 432 potential donors Reasons for excluding consenting Reasons for failure to collect Pregnancy related 63% donors prior to delivery Collection staff not at center or Personal medical history 12% unavailable 44% Pregnancy related 20% Personal risk behavior 8% Placenta/cord exclusion Personal medical history 41% Language barrier 7% 41% (including placental abruption) Personal risk behavior 20% Family history 3% Other delivery exclusion Family history 9% 7% Unable to consent 3% Other 10% Miscellaneous/undocumented 8% Private cord blood bank 2% Other / not documented 2% Figure 1 - Numbers of Women Screened, Approached, Consenting and with Collections Made Data from Monthly Recruitment Forms April 1998 to March 2001

9. Moved to long-term storage 9112 Total 64% 82% Cryopreserved collections Already shipped 303 11096 17207 Available in bank 8133 Still in quarantine 676 Reasons for discard 36% 18% Number Percent Insufficient volume/cell count 5171 64 Discarded Discarded Positive maternal ID test 646 8 6111 1984 Maternal history exclusion 613 8 Processing issues 297 4 Collection issues 294 4 No maternal sample 243 3 Subsequent medical information 224 3 Failed post-processing sterility test 194 2 Insufficient cell recovery 170 2 Withdrew consent 90 1 Administrative issues 57 1 Shipping/storage issues 91 1 Missing 5 0 Total 8095 100 Figure 2 - Numbers of Collections Made, Cryopreserved and Banked (December 1997 to August 2001)

10. Table 1 - Influence of Type of Delivery on Cell Counts and Volume Note : All differences highly significant (P< 0.0001).

11. Table 2 - Weight, Volume and Cell Counts of Stored CBUs by Ethnic Group

12. Table 3 - Percent of Collections Stored by Mother's Ethnicity

13. Table 4 - Correlations of Cell Counts, CBU Volume, Infant Birth Weight and Gestational Age *** P<0.001

14. 12 11 10 9 TNC (x 108) 8 7 <2.5 2.5 - 3.0 3.0 - 3.5 3.5 - 4.0 > 4.0 Birth weight (kg) Figure 2a - Total Nucleated Cells by Birth Weight

15. 5.0 4.5 4.0 CD34 (x 106) 3.5 3.0 2.5 2.0 <2.5 2.5 - 3.0 3.0 - 3.5 3.5 - 4.0 > 4.0 Birth weight (kg) Figure 2b - Total CD34 Cells by Birth Weight Bars show mean and 95% confidence limits (mean +/- 1.96 standard errors).

16. 12 11 10 TNC (x 108) 9 8 7 <37 38 39 40 > 40 Gestational Age (weeks) Figure 2c - Total Nucleated Cells by Gestational Age

17. 5.0 4.5 4.0 CD34 (x 106) 3.5 3.0 2.5 2.0 <37 38 39 40 > 40 Gestational Age (weeks) Figure 2d - Total CD34 Cells by Gestational Age

18. 3.0 2.5 Adjusted mean CD34+ (x 106) Total post processing count 2.0 1.5 Caucasian Hispanic Other African American Asian Figure 3 - Adjusted Mean Total Post-Processing CD34 Counts by Race

19. Conclusions • The COBLT Banking Study has demonstrated the resources required to establish and maintain a network of cord blood banks with common SOPs and Quality Systems. • A MCC (EMMES) created and maintains a web based data management system for data collection and analysis.

20. Conclusions • >35,000 women were recruited and screened over 3 years. • 15% refused to participate in the study. • Approximately 53% of CBUs collected from these donors were successfully banked creating an ethnically diverse inventory of >8,000 total units. • The main reasons for exclusion were insufficient cell volume/counts or positive ID testing on the maternal sample.

21. Conclusions • Strong correlations were observed between UCB collection volume and nucleated cell and CD34 content. • Units collected from African American donors had lower cell counts and CD34 counts per ml of UCB collected. • Factors correlating with higher UCB TNC content included higher birth weight, delivery by c-section and increasing gestational age at delivery (not beyond 40 weeks).

22. Conclusions • These observations can be used to create an algorithm for identification of desirable units at a collection site to maximize resources committed to units that are processed, tested and banked.

23. Acknowledgements • NHLBI: LeeAnn Jensen, Paul McCurdy • Study Chair: Nancy Kernan • The MCC – The EMMES Corporation • Shelly Carter, Liz Wagner, Geoff Cohen • Bank Directors • Mitch Cairo, John Fraser, Joanne Kurtzberg • HLA • UCSF, UCLA, NMRI