Stereological Quantification of Cell-Cycle Kinetics and Mobilization of Epithelial Stem Cells during Wound Healing
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We describe a stereology method to obtain reliable estimates of the total number of proliferative and migratory epithelial cells after wounding. Using pulse and chase experiments with halogenated thymidine analogs such as iododeoxyuridine (IdU) and chlorodeoxyuridine (CldU), it is possible to track epithelial populations with heterogeneous proliferative characteristics through skin compartments. The stereological and tissue processing methods described here apply widely to address important questions of skin stem-cell biology.
Key wordsStereology Hair follicles Halogenated thymidine analogs Iododeoxyuridine Chlorodeoxyuridine Bromodeoxyuridine Nucleoside Stem cells Bulge Epidermis Wound Keratinocytes Random sampling Confocal microscopy
EMM was a Research Fellow supported by Consejo Nacional de Ciencia y Tecnología (CONACYT; Estancias Postdoctorales y Sabáticas al extranjero: 175413) and Fundación México en Harvard, A.C. This work was supported by grants 38615N, 45872M, and 53194 from CONACYT, grant IN215208 from Dirección General de Asuntos del Personal Académico, UNAM (DGAPA), and grant IMPULSA-02 from Coordinación de la Investigación Científica, UNAM (CIC-UNAM) to GGO. Grant PAPIIT IA202013-2 from DGAPA, UNAM to EUQ. The authors are grateful to Ling Ling Lok for her careful editing.
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