title: Cell Cycle Analysis of CML Stem Cells Using Hoechst 33342 and Propidium Iodide.
abstract: Chronic myeloid leukemia (CML) is a myeloproliferative disease with an expansion of white blood cells. The current treatments for CML are shown not to be long-term effective because of CML stem cells' insensitivity to tyrosine kinase inhibitors. Therefore, studying more about CML stem cells is essential to understand the pathways of CML stem cell development and proliferation and finally lead to effective treatments to eliminate CML stem cells and eradicate CML. This chapter describes two methods to analyze cell cycle of CML stem cells. The rare population of CML stem cells can be identified by staining with cell surface markers, and then DNA-binding dyes Hoechst 33342 and propidium iodide (PI) are added to stain the DNA content which is changed when cells go through different phases of the cell cycle. Samples are run through the flow cytometer to be analyzed based on different absorbance and emission wavelengths of different florescent colors.
authors: University of Massachusetts Medical School, 364 Plantation Street, Worcester, MA, 01605, USA.
authors: Ngoc
authors: N
authors: DeSouza
authors: Acton-Boxborough Regional High School, 40 Lexington Drive, Acton, MA, 01720, USA.
authors: Megan
authors: M
authors: Zhou
authors: University of Massachusetts Medical School, 364 Plantation Street, Worcester, MA, 01605, USA. yi.shan@umassmed.edu.
authors: Yi
authors: Y
authors: Shan
mesh_terms: D000818:Animals; D001562:Benzimidazoles; D014408:Biomarkers, Tumor / Q000235:genetics*; D002453:Cell Cycle; D049109:Cell Proliferation; D004396:Coloring Agents; D004195:Disease Models, Animal; D005434:Flow Cytometry; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000235:genetics*; D051379:Mice; D014411:Neoplastic Stem Cells / Q000166:cytology*; D011419:Propidium
chemical_list: D001562:Benzimidazoles; D014408:Biomarkers, Tumor; D004396:Coloring Agents; D011419:Propidium; C017807:bisbenzimide ethoxide trihydrochloride
keywords: Cell cycle analysis; Chronic myeloid leukemia (CML) stem cells; Flow cytometry; Hoechst 33342; Ki67; Propidium iodide (PI)