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Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination John P....
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Transcript of Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination John P....
![Page 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,](https://reader035.fdocuments.us/reader035/viewer/2022062716/56649dc55503460f94ab8526/html5/thumbnails/1.jpg)
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
![Page 2: Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination John P. Miller, M.D., Ph.D. VP and Senior Medical Director,](https://reader035.fdocuments.us/reader035/viewer/2022062716/56649dc55503460f94ab8526/html5/thumbnails/2.jpg)
Background: Cord Blood Banking and Transplantation
• Recruitment: prenatal or at L&D suite
• Collection: in utero 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
![Page 3: Cord Blood Collection and Banking: Overview and Efforts to Minimize Microbial Contamination John P. Miller, M.D., Ph.D. VP and Senior Medical Director,](https://reader035.fdocuments.us/reader035/viewer/2022062716/56649dc55503460f94ab8526/html5/thumbnails/3.jpg)
Cord Blood Recruitment and Collection
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Collection Set-up and PreparationEx Utero
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Venipuncture and Completing the Collection
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Final Product
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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
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Cord blood receipt and inspection Rinse into transfer bag with HES
Inspection:Product intact, with no leaks clots or discoloration
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Soft spin to pellet RBCsExpress “buffy coat” from RBC pellet
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RBCs separated fromBuffy Coat and Plasma After Soft Spin
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Hard spin to pellet RBCs and WBCsExpress cell poor plasma from RBC pellet
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Transfer to Freezing Bag and Place in CBU Freezing Cassette
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Controlled Rate Freezing and Storage in LN2
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Potential Sources of Microbial Contamination of Cord Blood
• Cross-placental transmission• Contamination of cord or placenta during
delivery• Contamination during the collection
procedure
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Procedures to Minimize the Risk of Microbial Contamination
Organizational level
• NMDP Standards• NMDP Participation Criteria• NMDP membership process• AABB and FACT Standards and
accreditation
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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
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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
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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
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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
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Microbial contamination:NMDP banks through March 2004
33.0
11.3
11.5
17.28.5
6.2
4.8
4.8
2.8
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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
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Questions?
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