1 / 26

Cord Blood (CB) Hematopoietic Progenitor Cell (HPC) Characterization and Correlation with Ethnicity: A Report From The C

Cord Blood (CB) Hematopoietic Progenitor Cell (HPC) Characterization and Correlation with Ethnicity: A Report From The COBLT STUDY. 1 M S Cairo, 2 G Cohen, 2 E.L. Wagner, 3 J Fraser, 4 L Jensen, 2 S Carter, 5 N.A. Kernan, 6 J Kurtzberg

elam
Download Presentation

Cord Blood (CB) Hematopoietic Progenitor Cell (HPC) Characterization and Correlation with Ethnicity: A Report From The C

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Cord Blood (CB) Hematopoietic Progenitor Cell (HPC) Characterization and Correlation with Ethnicity: A Report From The COBLT STUDY 1M S Cairo, 2G Cohen, 2E.L. Wagner, 3J Fraser, 4L Jensen, 2S Carter, 5N.A. Kernan, 6J Kurtzberg 1Georgetown University, Washington DC, 2The Emmes Corporation, Rockville MD, 3University of California Los Angeles, Los Angeles CA, 4National Heart, Lung and Blood Institute, Bethesda MD, 5Memorial Sloan Kettering Cancer Center, New York NY, 6Duke University Medical Center, Durham NC

  2. Introduction • Advantages of CB and CBT Ease and safe procurement Enriched HPC Rapid availability Immaturity of T cell immunity Decreased viral transmission Decreased severe AGVHD Multi-ethnic representation Reduced chronic GVHD • Disadvantages of CB and CBT Decreased supply Delay in hematopoietic recovery Expensive to develop Prolonged immune reconstitution Limited cell dose Lack of available cells for ACI Genetic/infectious transmission Increased infectious morbidity

  3. Introduction • CB CD34+/kg graft content has been significantly correlated with engraftment and survival following UCBT Wagner et al, Blood, 2002 Styczynski et al, BMT, 2004 • CB vs BM CD34+/38- has a higher cloning efficiency, proliferate more rapidly and generate more HPC Cardoso et al, PNAS, 1993 Hao et al, Blood, 1995 • CB CD34+/CD90+ (Thy-1+) possess a high clonogenic cell expansion capacity and increased HPP-CFC Mayani et al, Blood, 1994 • PB CD34+/CD41+ (CD61+) subsets are significantly correlated with platelet recovery following Auto PBSCT Derecksen et al, JCO, 1998

  4. Objectives • To characterize processed CB HPC subsets including CD34+, CD34+/CD38-, CD34+/CD90+ and CD34+/CD61+ • To correlate processed CB HPC subsets with in vitro CB HPC colony formation including total CFU, CFU-GEMM, CFU-GM and BFU-E • To determine the relationship of birth weight, gestational age, ethnicity, gender and type of delivery with processed CB HPC

  5. Cord Blood Banks, Data Coordinating Center and NHLBI Cord Blood Bank Duke University J. Kurtzberg PI COBLT Study Committee N.A.Kernan, Chair/L Jensen, NHLBI Data Coordinating Center (Emmes) S. Carter & E.L. Wagner Cord Blood Bank Georgetown University M.Cairo, PI Cord Blood Bank University of California, LA J Fraser, PI

  6. Methods • Cord blood collection and processing were performed as described by COBLT Fraser,Cairo, Kurtzberg J. Hematotherapy, 1998 • Cord Blood CD34+ and CD34+ subset characterization (300-1200 events) • ProCount®/FACSort™ — Duke and UCLA • IgG1 FITC isotype control • ISHAGE/Epics XL-MCL —Georgetown University • IgG1 PE isotype control

  7. Methods * Duke & UCLA

  8. Methods • In vitro HPC Colony Assays • Processed CB (1 x 105 nucleated cells /mL) in IMDM, 2% FBS in Methocult® media • Incubated at 37°C, 5% for 11-14 days in 24 well tissue culture plates. • BFU-E • Bright red ≥ 50 cells/burst, tightly packed, multicentered • CFU-GM • Colorless, granular, ≥ 30 cells/colony, dispersed formation • CFU-GEMM • “Fried egg” appearance, ≥ 40 cells/colony, compact, spherical hemoglobinized area at center or one side.

  9. Results Maternal Donor Screened 34,799 (100%) Long Term Storage 8,731 53% of Collections 45% Pregnancy related 29% Refused 9% Medical History 6% Risk Behavior 11% Other Maternal Donor Consented 20,710 (60%) Low Counts Poor Viability Positive Culture No HPC Positive HGB Screen 44% No Staff 41% Exclusions 15% Other Cryopreserved 11,077 (66%) Total Collections 16,764 (81%) 64% CB Volume 11% +ID 7% History 18% Other

  10. Ethnic Distribution of Long Term Stored Cryopreserved UCB

  11. Delivery, Gender, Gestational Age, Birth Weight • Type of DeliveryGender • Vaginal 74% Males 52% • C-Section 26% Females 48% • Gestational Age (wks) Birth Weight (gms) • < 37 8% < 2500 1% • 38 17% 2501-3000 11% • 39 30% 3001-3500 37% • 40 31% 3501-4000 36% • 41 14% >4000 15%

  12. Cord Blood Volume, TNC, TMNC, CD34+

  13. Representative Flow Cytometry Dot Plots of CD34+, CD34+/CD38-, CD34+/CD61+ and CD34+/CD90+ CD34+ CD34+/CD38- CD34+/CD61+ CD34+/CD90+

  14. Cord Blood Hematopoietic Progenitor Cells (HPC)

  15. Log All CFU (x 104) Log CD34+ (x 104) Significant Correlation of CB Log CD34+ vs Log Total CFU

  16. Significant* Correlations of CB Log CD34+ vs CFU-GM, CFU-GEMM & BFU-E CD34+ vs CFU-GM CD34+ vs CFU-GEMM CD34+ vs BFU-E r = 0.68 P = 0.000 n = 8696 r = 0.52 p = 0.000 n = 4189 r = 0.61 p = 0.000 n = 8689 * All p<0.001

  17. Positive Correlation of Log Total Nucleated Cell Count (TNCC) with Log CD34+ Count in COBLT Processed Cord Blood

  18. Significant* Correlations of CB Log CD34+ subsets with Log CFUs * All p<0.001

  19. Significant* Correlations of CB Log CD34+ Subsets with Each Other * All p<0.0001

  20. 63 53 21 44 44 14 White African American Asian Hispanic Other White African American Asian Hispanic Other Other Other White White Black Black Asian Asian Hispanic Hispanic Significant* Reduction in CB CD34+/CD38- and CD34+/CD61+ in African Americans & Asians vs Caucasians & Hispanics CD34+/CD38- CD34+/CD61+ * All p<0.001

  21. Correlation of CB HPC with Birth Weight and Gestational Age Birth Weight Gestational Age CD34+/CD61+ CD34+/CD61+ CD34+/CD38- CD34+/CD38-

  22. Significant Correlation of CB HPC with Delivery and Gender

  23. Summary • Post processing CB log CD34+ was significantly correlated with log total CFU, BFU-E, CFU-GM and CFU-GEMM • Post processing CB log CD34+ subsets (CD38-, CD61+, CD90+) were significantly correlated with log total CFU, BFU-E, CFU-GM and CFU-GEMM • Post processing CB log CD34+ subsets were significantly correlated with each other (CD34+/CD38-, CD34+/CD61+, CD34+/CD90+)

  24. Summary • African-American and Asian donors were associated with significantly less CD34+/CD38- and CD34+/CD61+ processed CB HPC subsets • C-section delivery was significantly associated with increased total processed CB CFU • Male gender was significantly associated with increased CB CD34+/CD61+ HPC subset • Birth weight was significantly correlated with both CD34+/CD38- and CD34+/CD61+ and inversely correlated with gestational age.

  25. Speculation • Post processing CB HPC CD34+ subsets and/or in vitro CB HPC CFUs may potentially correlate with the kinetics of lineage specific CB engraftment post CBT • Ethnicity, gender, delivery, birth weight and /or gestational age may have a significant effect on CB HPC and therefore, may be a factor in the future selection of a processed CB unit for CBT • The current COBLT UCBT study will determine the significance of these observations.

  26. Acknowledgments • NHLBI Support • N01-HB-67136, N01-HB-67142, N01-HB-67132 and N01-HB-67141 • Cord Blood collection, processing and flow cytometry personnel at Carolina, UCLA and Georgetown University Cord Blood Banks • Maternal donors and newborns for their public generosity

More Related