A novel method for efficient generation of transfected human osteoclasts.
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 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.
Taylor
A
A
Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK. adam.taylor@abdn.ac.uk
Rogers
M J
MJ
Tosh
D
D
Coxon
F P
FP
D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D002470:Cell Survival; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods; D005818:Genetic Engineering; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D053245:RANK Ligand / Q000494:pharmacology; D014162:Transfection / Q000379:methods*
D053245:RANK Ligand; D049452:Green Fluorescent Proteins