Novel application of a fibrin cell block method to evaluate melanocytic cell populations
| Autor: | Lester J. Layfield, Courtney B. McKinney, Kenneth M. Boucher, Scott R. Florell, Patricia Porter-Gill, Diana L. Biddle, Douglas Grossman, Soo J. Schmidt, Kurt H. Albertine, Frederic Clayton, Sancy A. Leachman, Kelley J. Murphy |
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| Rok vydání: | 2003 |
| Předmět: |
Adult
Clinical Biochemistry Cell Population Antigens Differentiation Myelomonocytic Plant Science Melanocyte Fibrin Flow cytometry MART-1 Antigen Antigens CD Antigens Neoplasm medicine Humans education Cells Cultured Skin education.field_of_study medicine.diagnostic_test biology Cell Biology Fibroblasts Flow Cytometry Molecular biology Immunohistochemistry Staining Neoplasm Proteins HMB-45 medicine.anatomical_structure Cell culture Immunology biology.protein Melanocytes Agronomy and Crop Science Melanoma-Specific Antigens Developmental Biology |
| Zdroj: | Pigment cell research. 16(6) |
| ISSN: | 0893-5785 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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