title: Efficient transient transfection of human multiple myeloma cells by electroporation--an appraisal.
abstract: Cell lines represent the everyday workhorses for in vitro research on multiple myeloma (MM) and are regularly employed in all aspects of molecular and pharmacological investigations. Although loss-of-function studies using RNA interference in MM cell lines depend on successful knockdown, no well-established and widely applied protocol for efficient transient transfection has so far emerged. Here, we provide an appraisal of electroporation as a means to introduce either short-hairpin RNA expression vectors or synthesised siRNAs into MM cells. We found that electroporation using siRNAs was much more efficient than previously anticipated on the basis of transfection efficiencies deduced from EGFP-expression off protein expression vectors. Such knowledge can even confidently be exploited in "hard-to-transfect" MM cell lines to generate large numbers of transient knockdown phenotype MM cells. In addition, special attention was given to developing a protocol that provides easy implementation, good reproducibility and manageable experimental costs.
authors.affiliation: Department of Internal Medicine II, Division of Hematology and Oncology, University Hospital of Würzburg, Würzburg, Germany.
authors.forename: Torsten
authors.initials: T
authors.lastname: Steinbrunn
authors.affiliation: Department of Internal Medicine II, Division of Hematology and Oncology, University Hospital of Würzburg, Würzburg, Germany.
authors.forename: Manik
authors.initials: M
authors.lastname: Chatterjee
authors.affiliation: Comprehensive Cancer Center Mainfranken, University Hospital of Würzburg, Würzburg, Germany.
authors.forename: Ralf C
authors.initials: RC
authors.lastname: Bargou
authors.affiliation: Department of Internal Medicine II, Division of Hematology and Oncology, University Hospital of Würzburg, Würzburg, Germany.
authors.forename: Thorsten
authors.initials: T
authors.lastname: Stühmer
mesh_terms: D045744:Cell Line, Tumor; D002470:Cell Survival; D018274:Electroporation / Q000379:methods*; D005434:Flow Cytometry; D015870:Gene Expression; D055785:Gene Knockdown Techniques; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000235:genetics / Q000378:metabolism; D009101:Multiple Myeloma; D034622:RNA Interference; D034741:RNA, Small Interfering; D014162:Transfection / Q000379:methods*
chemical_list: D034741:RNA, Small Interfering; D019950:Mitogen-Activated Protein Kinase 1