Molecular characterisation of post-bio-electrosprayed human brain astrocytoma cells.
Bio-electrospraying (BES) is a method for directly jetting living cells under conditions that allow their distribution in the x, y, and z axes. Previous work has been focused on achieving jetting in stable cone-jet mode, which is required for precision placement, and these studies have demonstrated that there are no significant effects of bio-electrospraying on cell morphology or viability. In this work, we examine the biological properties of bio-electrosprayed cells using assays of cellular function that range from the molecular level through to integrated cellular systems, and include proteomics, signal transduction, cell growth and proliferation, and the characterisation of apoptotic blebs. From these molecular methods, we have determined that bio-electrospraying, under the electric field conditions used to achieve stable cone-jet mode, causes no alterations to the biological properties and function of the cells being jetted. Bio-electrosprayed and control cells had similar viability, proliferation properties and virtually indistinguishable cell cycle profiles. The biophysical properties of large conducting (BK) potassium channels were unchanged, as were the pharmacological responses of the endogenous muscarinic and exogenous P2Y(11) receptors, both of which are cell surface receptors of the 7TM superfamily. Proteomic analyses revealed that although three proteins had subtle differences in expression level between bio-electrosprayed and control cells, none of these fold differences was above the 1.5-fold cut-off threshold required for further analyses. These findings support the further development of bio-electrosprays as a viable technology for a wide diversity of tissue engineering, regenerative biology, advanced cellular therapeutics and medicinal applications, having significance in the clinic.
Eddaoudi
Ayad
A
Flow Cytometry Core Facility, Camelia Botnar Laboratories, UCL Institute of Child Health/Great Ormond Street Hospital for Children, 30 Guilford Street, London, WC1N 1EH, United Kingdom.
Townsend-Nicholson
Andrea
A
Timms
John F
JF
Schorge
Stephanie
S
Jayasinghe
Suwan N
SN
D001254:Astrocytoma; D001932:Brain Neoplasms; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D007211:Indoles / Q000494:pharmacology; D008301:Maleimides / Q000494:pharmacology; D015221:Potassium Channels / Q000378:metabolism; D043587:Receptor, Muscarinic M3 / Q000235:genetics / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D007211:Indoles; D008301:Maleimides; D015221:Potassium Channels; D043587:Receptor, Muscarinic M3; D014409:Tumor Necrosis Factor-alpha; D002118:Calcium; C064755:Ro 31-7549