Advertisement

Colony Formation: An Assay of Hematopoietic Progenitor Cells

  • Romy Kronstein-Wiedemann
  • Torsten TonnEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2017)

Abstract

The colony-forming cell (CFC) assay is used to study the proliferation and differentiation pattern of each input hematopoietic progenitors by their ability to form colonies in a semisolid medium. The resulting colonies are consisting of more differentiated cells, and the number and the morphology of the colonies provide preliminary information about the ability of progenitors to differentiate and proliferate. To allow colonies to grow to a size which facilitates accurate counting and identification, about 14 days of culture is sufficient. In certain situations also shorter periods may be used.

Key words

Ficoll Red blood cell depletion Hematopoietic progenitors Methyl cellulose Semisolid culture Colony-forming units Erythroid and myeloid colonies STEMvision 

References

  1. 1.
    Kosan C, Godmann M (2016) Genetic and epigenetic mechanisms that maintain hematopoietic stem cell function. Stem Cells Int 2016:5178965CrossRefGoogle Scholar
  2. 2.
    Barrett D, Fish JD, Grupp SA (2010) Autologous and allogeneic cellular therapies for high-risk pediatric solid tumors. Pediatr Clin N Am 57(1):47–66CrossRefGoogle Scholar
  3. 3.
    Milanetti F, Abinun M, Voltarelli JC, Burt RK (2010) Autologous hematopoietic stem cell transplantation for childhood autoimmune disease. Pediatr Clin N Am 57(1):239–271CrossRefGoogle Scholar
  4. 4.
    Özgüner M, Tavil B, Köksal J, Canal E, Bozkaya I, Tunç B (2011) Comparison of colony forming unit-assay results of different hematopoietic stem cell sources. Turkish J Pediatr Dis 5(4):197–201Google Scholar
  5. 5.
    Chervenick PA, Boggs DR (1971) In vitro growth of granulocytic and mononuclear cell colonies from blood of normal individuals. Blood 37(2):131–135PubMedGoogle Scholar
  6. 6.
    Clarke BJ, Housman D (1977) Characterization of an erythroid precursor cell of high proliferative capacity in normal human peripheral blood. Proc Natl Acad Sci U S A 74(3):1105–1109CrossRefGoogle Scholar
  7. 7.
    McCredie KB, Hersh EM, Freireich EJ (1971) Cells capable of colony formation in the peripheral blood of man. Science 171:293–294CrossRefGoogle Scholar
  8. 8.
    Juttner CA, To LB, Ho JQ, Bardy PG, Dyson PG, Haylock DN, Kimber RJ (1988) Early lympho-hemopoietic recovery after autografting using peripheral blood stem cells in acute non-lymphoblastic leukemia. Transplant Proc 20(1):40–42PubMedGoogle Scholar
  9. 9.
    Kessinger A, Armitage JO, Landmark JD, Smith DM, Weisenburger DD (1988) Autologous peripheral hematopoietic stem cell transplantation restores hematopoietic function following marrow ablative therapy. Blood 71(3):723–727PubMedGoogle Scholar
  10. 10.
    Korbling M, Dorken B, Ho AD, Pezzutto A, Hunstein W, Fliedner TM (1986) Autologous transplantation of blood-derived hemopoietic stem cells after myeloablative therapy in a patient with Burkitt’s lymphoma. Blood 67(2):529–532PubMedGoogle Scholar
  11. 11.
    Reiffers J, Bernard P, David B, Vezon G, Sarrat A, Marit G, Moulinier J, Broustet A (1986) Successful autologous transplantation with peripheral blood hemopoietic cells in a patient with acute leukemia. Exp Hematol 14(4):312–315PubMedGoogle Scholar
  12. 12.
    Udomsakdi C, Lansdorp PM, Hogge DE, Reid DS, Eaves AC, Eaves CJ (1992) Characterization of primitive hematopoietic cells in normal human peripheral blood. Blood 80(10):2513–2521PubMedGoogle Scholar
  13. 13.
    Eaves C (1995) Assays of hematopoietic progenitor cells. In: Beutler E, Lichtman MA, Coller BS, Kipps TJ (eds) Williams hematology, 5th edn. McGraw-Hill, New York, pp 22–66Google Scholar
  14. 14.
    Klug CA, Jordan CT (eds) (2002) Hematopoietic stem cell protocols, vol 63. Humana Press, Totowa, NJGoogle Scholar
  15. 15.
    Miller CL, Lai B (2005) Human and mouse hematopoietic colony-forming cell assays. Methods Mol Biol 290:71–89PubMedGoogle Scholar
  16. 16.
    Gordon MY (1993) Human haemopoietic stem cell assays. Blood Rev 7(3):190–197CrossRefGoogle Scholar
  17. 17.
    Eaves CJ (2015) Hematopoietic stem cells: concepts, definitions, and the new reality. Blood 125(17):2605–2613CrossRefGoogle Scholar
  18. 18.
    Low WS, Wan Abas WA (2015) Benchtop technologies for circulating tumor cells separation based on biophysical properties. Biomed Res Int 2015:239362PubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Institute for Transfusion Medicine, German Red Cross Blood Donation Service North-East, Medical Faculty Carl Gustav CarusTechnical University DresdenDresdenGermany

Personalised recommendations