Abstract
All stem cells exhibit the capacity and ability to proliferate. This is a fundamental property of stem cells, since it is required not only for self-renewal, but also for expansion and the ability for stem cells to engraft in a patient. The capacity or potential for proliferation by stem cells defines their degree of primitives or stemness. This, in turn, is directly related to stem cell self-renewal. Using a highly sensitive, accurate, and reliable ATP bioluminescence signal detection system that can be multiplexed with other assay readouts, it has been possible to determine three stem cell parameters using a single assay. Using primitive hematopoietic bone marrow stem cells as an example, an in vitro protocol is described that incorporates initially culturing primitive stem cells to induce them into cell cycle, followed by a secondary re-plating step that demonstrates both self-renewal capability and expansion potential.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Bradley TR, Metcalf D (1966) The growth of mouse bone marrow cells in vitro. Aust J Exp Biol Med Sci 44:287–299
Pluznik DH, Sachs L (1966) The induction of clones of normal mast cells by a substance from conditioned medium. Exp Cell Res 43:553–563
Iscove NN, Sieber F, Winterhalter KH (1974) Erythroid colony formation in cultures of mouse and human bone marrow: analysis of the requirement for erythropoietin by gel filtration and affinity chromatography on agarose-concanavalin A. J Cell Physiol 83:309–320
McQualter JL, Yuen K, Williams B et al (2010) Evidence of an epithelial stem/progenitor cell hierarchy in the adult mouse lung. Proc Natl Acad Sci U S A 107:1414–1419
Povsic TJ, Zavodni KL, Vainorius E et al (2009) Common endothelial progenitor cell assays identify discrete endothelial progenitor cell populations. Am Heart J 157:335–344
Zhang L, Valdez JM, Zhang B et al (2011) ROCK inhibitor Y-27632 suppresses dissociation-induced apoptosis of murine prostate stem/progenitor cells and increases their cloning efficiency. PLoS One 6:e18271. doi:10.1371/journal.pone.0018271
Rosendaal M, Hodgson GS, Bradley TR (1979) Organization of haemopoietic stem cells: the generation-age hypothesis. Cell Tissue Kinet 12:7–29
Hodgson GS, Bradley TR (1979) Properties of haematopoietic stem cells surviving 5-fluorouracil treatment: evidence for a pre-CFU-S cell? Nature 381–2
Rich IN, Hall KM (2005) Validation and development of a predictive paradigm for hemotoxicology using a multifunctional bioluminescence colony-forming proliferation assay. Toxicol Sci 87:427–441
Hall KM, Harper H, Rich IN (2012) Hematopoietic stem cell potency for cellular therapeutic transplantation. In: Pelayo R (Ed.) Advances in hematopoietic stem cell research. InTech. ISBN: 978-953-307-930-1
Rich IN (2013) Potency, proliferation and engraftment potential of stem cell therapeutics: The relationship between potency and clinical outcome for hematopoietic stem cell products. J Cell Sci Ther. S13-001. doi:10.4172/2157-7013
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2015 Springer Science+Business Media New York
About this protocol
Cite this protocol
Rich, I.N. (2015). Measurement of Hematopoietic Stem Cell Proliferation, Self-Renewal, and Expansion Potential. In: Rich, I. (eds) Stem Cell Protocols. Methods in Molecular Biology, vol 1235. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1785-3_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-1785-3_2
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-1784-6
Online ISBN: 978-1-4939-1785-3
eBook Packages: Springer Protocols