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Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination

Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination. John P. Miller, M.D., Ph.D. VP and Senior Medical Director, NMDP May, 2009. Background: Cord Blood Banking and Transplantation. Recruitment: prenatal or at L&D suite

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Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination

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  1. Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination John P. Miller, M.D., Ph.D. VP and Senior Medical Director, NMDP May, 2009

  2. Background: Cord Blood Banking and Transplantation • Recruitment: prenatal or at L&D suite • Collection: inutero or ex utero • Processing: manual or automated plasma depletion +/- RBC depletion • Cryopreservation: 6%HES/10% DMSO • Storage: liquid or vapor phase of LN2 • Transport: Dry Shipper -196 to -150 C • Thawing: with or without washing • Infusion: ASAP

  3. Cord Blood Recruitment and Collection

  4. Collection Set-up and PreparationEx Utero

  5. Venipuncture and Completing the Collection

  6. Final Product

  7. Steps in cord blood processing • Receipt, inspection • Initial sampling: TNC, ABO/Rh • Centrifuge soft spin: sedimentation of RBCs with HES • Hard spin to concentrate HPCs • Product analysis: TNC, viability, culture, CD34, CFU • Cryopreservation • Distribution/shipping • Thawing and washing • Infusing

  8. Cord blood receipt and inspection Rinse into transfer bag with HES Inspection: Product intact, with no leaks clots or discoloration

  9. Soft spin to pellet RBCsExpress “buffy coat” from RBC pellet

  10. RBCs separated fromBuffy Coat and Plasma After Soft Spin

  11. Hard spin to pellet RBCs and WBCsExpress cell poor plasma from RBC pellet

  12. Transfer to Freezing Bag and Place in CBU Freezing Cassette

  13. Controlled Rate Freezing and Storage in LN2

  14. Potential Sources of Microbial Contamination of Cord Blood • Cross-placental transmission • Contamination of cord or placenta during delivery • Contamination during the collection procedure

  15. Procedures to Minimize the Risk of Microbial Contamination Organizational level • NMDP Standards • NMDP Participation Criteria • NMDP membership process • AABB and FACT Standards and accreditation

  16. Procedures to Minimize the Risk of Microbial Contamination At the CBU level: • Maternal health history screening for RCD • Maternal IDM testing for RCD • Review maternal prenatal and delivery history/exam for risk of transmissible disease, e.g. chorioamnionitis, sepsis • Review of infant history and exam • Examination of cord and placenta, e.g. tears, infection • Preparation of cord for venipuncture • Aseptic Processing- one unit at a time • Time from delivery to cryopreservation • Bacterial and fungal cultures of final product prior to cryopreservation • Post-discharge infant follow-up (bact and genetic) • Adverse event reporting and investigation

  17. Procedures to Minimize the Risk of Microbial Contamination Specific to Neisseria and Chlamydia: HHQ Donor is eligible with history of treatment for chlamydia for either type of delivery

  18. Microbial contamination:Literature Review • Contamination of CBU occurs at 2-5%1-3 • Culture Method: BacT-Alert or Bactec,Culture NOS organisms include: • Staphylococcus sp. • Streptococcus sp., including enterococcus • Corynebacterium sp. • E. coli • Bacteroides fragilis • No Neisseria gonorrhea or Chlamydia trachomatis • 1Bertolini 1995, 2M-Reboredo 2000,3Donaldson 2000

  19. Microbial contamination:NMDP banks through March 2004 • 356 CBUs had positive cultures (0.7%, n=51,842) • Culture Method: BacT-Alert or Bactec • Common organisms include: • Staphylococcus sp. • Streptococcus sp., including enterococcus • Corynebacterium sp. • Bacillus sp. • E. coli • Bacteroides fragilis • No ID Aerobes and Anaerobes • Mixed flora • No Neisseria gonorrhea or Chlamydia trachomatis

  20. Microbial contamination:NMDP banks through March 2004 6.2 11.3 11.5 33.0 4.8 2.8 4.8 8.5 17.2

  21. Summary • Steps to minimize bacterial contamination of CBUs include maternal and infant screening, IDM testing, examination of the cord and placenta, preparation for venipuncture, aseptic processing and culture • Rates of cord blood bacterial contamination are lower than reported in the literature (about 0.7%) • No cases of contamination with Neisseria or Chlamydia have been reported

  22. Questions? • ?????

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