Biomedical Engineering Reference
In-Depth Information
Chapter 6
The Cluster-Forming Activity Assay:
A Short-Term In Vitro Method
to Analyze the Activity of Mouse
Spermatogonial Stem Cells
Makoto C. Nagano and Jonathan R. Yeh
Abstract The previous two chapters discussed contrasting approaches to detect
SSCs and investigate their biology. The approach described in Chap. 4 represents
the attempt to identify SSCs prospectively by means of cell morphology, while the
transplantation approach in Chap. 5 is based on the functional definition of stem
cells (long-term self-renewal and differentiation) and represents retrospective SSC
identification. This chapter will discuss another type of retrospective, functional
SSC detection method, termed the “cluster-forming activity (CFA) assay.” This
technique was developed in the mouse model on the basis of the SSC culture sys-
tem described in Chap. 5. Using this in vitro assay, SSC activity can be detected
in a semi-quantitative manner within a short period of time, in marked contrast
to the time-consuming and laborious transplantation assay. As with any technol-
ogy, however, the CFA assay is not without limitations, and there are issues to be
noted when one uses it and interprets the data obtained. The aim of this chapter is
therefore twofold. First, we describe the conceptual framework of the CFA assay
in order to justify its legitimacy as a reliable SSC detection method. Second, we
discuss cautionary issues and relate them to the in vitro behavior of SSCs warranting
further studies.
Keywords Quantitative stem cell assay • Cell culture • Cluster • Colony
• Transplantation
M.C. Nagano ( * )
Department of Obstetrics and Gynecology and Division of Experimental Medicine,
McGill University, Royal Victoria Hospital, Room F3.07, 687 Pine Avenue West,
Montreal, Quebec H3A 1A1, Canada
e-mail: makoto.nagano@muhc.mcgill.ca
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