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

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

• 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

Cord Blood Recruitment and Collection

Collection Set-up and PreparationEx Utero

Venipuncture and Completing the Collection

Final Product

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

Cord blood receipt and inspection Rinse into transfer bag with HES

Inspection:Product intact, with no leaks clots or discoloration

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

RBCs separated fromBuffy Coat and Plasma After Soft Spin

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

Transfer to Freezing Bag and Place in CBU Freezing Cassette

Controlled Rate Freezing and Storage in LN2

Potential Sources of Microbial Contamination of Cord Blood

• Cross-placental transmission• Contamination of cord or placenta during

delivery• Contamination during the collection

procedure

Procedures to Minimize the Risk of Microbial Contamination

Organizational level

• NMDP Standards• NMDP Participation Criteria• NMDP membership process• AABB and FACT Standards and

accreditation

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

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

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

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

Microbial contamination:NMDP banks through March 2004

33.0

11.3

11.5

17.28.5

6.2

4.8

4.8

2.8

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

Questions?

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