Quantitative accounting of CD34+ hematopoietic stem cells in whole cord blood

Cover Page


Cite item

Full Text

Open Access Open Access
Restricted Access Access granted
Restricted Access Subscription or Fee Access

Abstract

Umbilical cord blood is an alternative source of hematopoietic stem cells to bone marrow and mobilized peripheral blood [HSC]. The population of CD34" cells in umbilical cord blood is small, which creates certain difficulties in carrying out quantitative accounting of HSC. This article discusses some practical aspects of determining CD34" HSC in whole cord blood. A preliminary assessment of the amount of HSC in the samples before the start of preparation for long-term storage may become an additional selection criterion when receiving cord blood in non-commercial anonymous banks.

Full Text

Restricted Access

About the authors

A. V. Kurtova

Laboratory Diagnostics Center of St. Petersburg State Medical University named after Academician IL. Pavlov

Author for correspondence.
Email: redaktor@celltranspl.ru

Laboratory of Clinical Immunology and Molecular Diagnostics

Russian Federation, Saint-Petersburg

E. E. Zueva

Laboratory Diagnostics Center of St. Petersburg State Medical University named after Academician IL. Pavlov

Email: redaktor@celltranspl.ru

Laboratory of Clinical Immunology and Molecular Diagnostics

Russian Federation, Saint-Petersburg

References

  1. Watt S., Contreras М. Stem cell medicine: Umbilical cord blood and its stem cell potential. Semin. Fetal. Neonat. Med. 2005; 10: 209-20.
  2. Bradley M.B., Cairo M.S. Cord blood immunology and stem cell transplantation. Human Immunol. 2005; 6: 431-46.
  3. International Standards for Cord Blood Collection, Processing, Testing, Banking, Selection and Release. NETCORD/FACT 2006; 1-65.
  4. Warwick R., Armitage S. Cord blood banking. Best Pract. Res. Clin. Obstet. Gynaecol. 2004; 18(6): 995-1011.
  5. Gluckman E., Rocha V., Chastang C. Use of cord blood cells for banking and transplant. The Oncologist 1997; 2: 340-3.
  6. Tse W„ Laughlin M.J. Umbilical cord blood transplantation: a new alternative option. Hematology Education Programme Book 2005; 377-83.
  7. Nakagawa R., Watanabe T„ Kawano Y. et al. Analysis of maternal and neonatal factors that influence the nucleated and CD34+ cell yield for cord blood banking. Transfusion 2004; 44[2): 262-7.
  8. Gratama J., Sutherland D.R., Keeney M„ Papa S. Flow cytometric enumeration and immunophenotyping of hematopoietic stem and progenitor cells. J. Biol. Regul. Homeost. Agents. 2001; 15:14-22.
  9. Sutherland D.R., Anderson L, Keeney M. et al. The ISHAGE guidelines for CD34+ cell determination by flow cytometry. J. Hematother. 1996; 5:213-26.
  10. Gutensohn K., Serke S., Cassens U. et al. German sociétés of hematology/oncology and transfusion medicine and immunohematology for flow cytometric CD34-determination. Infusionsther Transfusionmed. 1996; 23[suppl 2): 1-23.
  11. Brocklebank A.M., Sparrow R.L. Enumeration of CD34+ cells in cord blood: a variation on a single-plantform flow cytometric method based on the ISHAGE gating strategy. Cytometry 2001; 46: 254-61.
  12. Зуева E.E. Молчанова И.В., Комарова Л.С. и др. Особенности количественного определения гемопоэтических стволовых клеток методом проточной цитометрии. Молекулярная медицина 2006; В печати.
  13. Laroche V., McKenna D.H., Moroff G. et al. Cell loss and recovery in umbilical cord processing: a comparison of postthaw and postwash samples. Transfusion 2005; 45:1909-16.

Supplementary files

Supplementary Files
Action
1. JATS XML
2. Fig. 1. Two-dimensional histogram of light scattering parameters of a sample of whole cord blood. The unusual pattern of distribution of populations of lymphocytes and monocytes draws attention. The affiliation of the clusters to these populations was determined by analyzing the CD45/SSC histogram

Download (53KB)
3. Fig. 2. The sequence of creating logical constraints during the quantitative accounting of GSK

Download (174KB)
4. Fig. 3. Sample N. Two-dimensional histogram of light scattering parameters. Imposing a logical restriction on leukocytes on the CD45/CD34 histogram in this case does not allow unambiguously identifying the HSC population.

Download (155KB)
5. Fig. 4. Sample N. Two-dimensional histogram of light scattering parameters. The imposition of a logical restriction on mononuclears on the CD45/CD34 histogram makes it possible to identify the target population, which is confirmed by the quantitative coincidence of HSCs in gates R2 and R4

Download (81KB)
6. Fig. 5. Identification of two populations of CD34+ cells in a sample of whole cord blood prepared according to the staining-lysis protocol. Reverse gating confirmed that both populations belong to the lymphocyte region

Download (130KB)
7. Fig. 6. A sample of whole cord blood with a very low content CD34+ HSC cells are small in number and do not form clusters, which indicates the absence of clonogenic activity in them

Download (98KB)

Copyright (c) 2023 Eco-Vector



СМИ зарегистрировано Федеральной службой по надзору в сфере связи, информационных технологий и массовых коммуникаций (Роскомнадзор).
Регистрационный номер и дата принятия решения о регистрации СМИ: 

This website uses cookies

You consent to our cookies if you continue to use our website.

About Cookies