Novel Application of a Fibrin Cell Block Method to Evaluate Melanocytic Cell Populations

Scott R. Florell, Soo J. Schmidt, Patricia Porter-Gill, Kurt H. Albertine, Kelley J. Murphy, Courtney B. McKinney, Kenneth M. Boucher, Douglas Grossman, Diana L. Biddle, Frederic Clayton, Lester J. Layfield, Sancy A. Leachman

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Confirming melanocytic lineage and purity is important for experiments using cultured human melanocytes. The objective of this study was to develop a simple, reliable method to evaluate and archive cultured melanocytic cells. Melanocytes were isolated from adult skin biopsies or from neonatal foreskins using standard culturing methods. Fibrin cell blocks (FCBs) were prepared from cultured cells at passages two and six. Fibrin blocks were paraffin-embedded and sectioned for immunohistochemical (CD68, Melan-A, and HMB-45) and H & E staining. Flow cytometry was performed (Melan-A) at passage six. A mixing experiment with cultured melanocytes and fibroblasts was performed and cell population purity was determined by manual counts of positively staining cells in the FCBs and by flow cytometry. The FCB method of evaluating population purity was validated experimentally and by correlation with flow cytometry results. Preparation of a FCB followed by immunohistochemical staining is an easy and inexpensive way to confirm melanocytic lineage, estimate population purity, and provide a permanent archive of cultured cells.

Original languageEnglish (US)
Pages (from-to)662-669
Number of pages8
JournalPigment Cell Research
Volume16
Issue number6
DOIs
StatePublished - Dec 1 2003

Keywords

  • Cell block
  • Cell culture
  • Culture
  • Fibrin
  • Formalin
  • HMB-45
  • Melan-A
  • Melanocyte
  • Paraffin

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science
  • Developmental Biology
  • Clinical Biochemistry
  • Cell Biology

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  • Cite this

    Florell, S. R., Schmidt, S. J., Porter-Gill, P., Albertine, K. H., Murphy, K. J., McKinney, C. B., Boucher, K. M., Grossman, D., Biddle, D. L., Clayton, F., Layfield, L. J., & Leachman, S. A. (2003). Novel Application of a Fibrin Cell Block Method to Evaluate Melanocytic Cell Populations. Pigment Cell Research, 16(6), 662-669. https://doi.org/10.1046/j.1600-0749.2003.00095.x