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Stereological Quantification of Cell-Cycle Kinetics and Mobilization of Epithelial Stem Cells during Wound Healing

  • Eduardo Martínez-MartínezEmail author
  • Eileen Uribe-Querol
  • Claudio I. Galván-Hernández
  • Gabriel Gutiérrez-Ospina
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1453)

Abstract

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 words

Stereology Hair follicles Halogenated thymidine analogs Iododeoxyuridine Chlorodeoxyuridine Bromodeoxyuridine Nucleoside Stem cells Bulge Epidermis Wound Keratinocytes Random sampling Confocal microscopy 

Notes

Acknowledgements

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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Eduardo Martínez-Martínez
    • 1
    • 2
    • 3
    Email author
  • Eileen Uribe-Querol
    • 4
  • Claudio I. Galván-Hernández
    • 3
  • Gabriel Gutiérrez-Ospina
    • 3
  1. 1.Laboratory of RNA & Extracellular VesiclesInstituto Nacional de Medicina GenómicaTlalpanMéxico
  2. 2.Department of MedicineBrigham and Women’s Hospital, Harvard Medical SchoolBostonUSA
  3. 3.Departamento de Biología Celular y Fisiología and Grupo Multidisciplinario de Investigación en Células Troncales IMPULSA 02Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de MéxicoCoyoacánMexico
  4. 4.División de Estudios de Posgrado e InvestigaciónFacultad de Odontología, Universidad Nacional Autónoma deMéxicoCoyoacánMexico

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