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Detection of apoptotic cells in vitro: a practical standardized experimental protocol using human fibroblasts for
H E Gruber1, S S Coleman, A A Stasky
1Department of Orthopaedic Surgery, Carolinas HealthCare System, Charlotte, North Carolina 28232, USA. hgruber@carolinas.org
Abstract:
This technical report presents a practical experimental protocol using human fibroblasts that is useful for initiating in situ methods for detection of apoptotic cells in vitro. The protocol details the growth of cells in multichamber wells, and includes a negative control, two positive controls, and untreated fibroblasts to be evaluated for apoptosis localization. Technical tips on handling of solutions, cell culture designs, and separation of slides by treatment are valuable steps for the laboratory beginning such studies. The methods proposed are both time and cost effective.