A novel method for efficient generation of transfected human osteoclasts

Authors:
Taylor A, Rogers MJ, Tosh D, Coxon FP
In:
Source: Calcif Tissue Int
Publication Date: (2007)
Issue: 80(2): 132-6
Research Area:
Immunotherapy / Hematology
Cells used in publication:
Osteoclast precursor (OCP), human
Species: human
Tissue Origin: bone marrow
Platform:
Nucleofector® I/II/2b
Experiment
"The Amaxa nucleofection procedure resulted in successful tranfection of human osteoclast precursors..." ; "This technique offers an attractive alternative to adenoviral transfeciton since it has the advantages of being very simple (requiring only plasmid DNA) and extremely quick."
Abstract
Mature osteoclasts and their precursors are notoriously difficult to transfect using nonviral approaches, a limitation that represents a major technical obstacle in the study of osteoclast biology. Here, we describe a simple electroporation method using Amaxa((R)) Nucleofector technology that results in efficient transfection of human blood-derived osteoclast precursors, which can be differentiated in subsequent culture to generate mature osteoclasts that retain expression of the transgene. Moreover, since these osteoclasts maintain the ability to resorb dentine, this technique could prove useful for assessing the role of specific genes/proteins in osteoclast function.