Collision of millimetre droplets induces DNA and protein transfection into cells

نویسندگان

  • Kazuto Ikemoto
  • Ichiro Sakata
  • Takafumi Sakai
چکیده

Nonperturbing and simple transfection methods are important for modern techniques used in biotechnology. Recently, we reported that electrospraying can be applied to DNA transfection in cell lines, bacteria, and chicken embryos. However, the transfection efficiency was only about 2%. To improve the transfection rate, physical properties of the sprayed droplets were studied in different variations of the method. We describe a highly efficient technique (30-93%) for introduction of materials such as DNA and protein into living cells by electrospraying droplets of a high conductivity liquid onto cells incubated with the material for transfection. Electric conductivity has a sizable influence on the success of transfection. In contrast, molecular weight of the transfected material, types of ions in the electrospray solution, and the osmotic pressure do not influence transfection efficiency. The physical analysis revealed that collision of cells with millimetre-sized droplets activates intracellular uptake.

برای دانلود رایگان متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Optimization of SW480 Colon Cancer Cells Transfection with Lipofectamine 2000

ABSTRACT           Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transg...

متن کامل

Downregulation of Kinesin Spindle Protein Inhibits Proliferation, Induces Apoptosis and Increases Chemosensitivity in Hepatocellular Carcinoma Cells

Background: Kinesin spindle protein (KSP) plays a critical role in mitosis. Inhibition of KSP function leads to cell cycle arrest at mitosis and ultimately to cell death. The aim of this study was to suppress KSP expression by specific small-interfering RNA (siRNA) in Hep3B cells and evaluate its anti-tumor activity. Methods: Three siRNA targeting KSP (KSP-siRNA #1-3) and one mismatched-siRNA (...

متن کامل

Highly Efficient Transfection of Dendritic Cells Derived from Esophageal Squamous Cell Carcinoma Patient: Optimization with Green Fluorescent Protein and Validation with Tumor RNA as a Tool for Immuno-genetherapy

This study was conducted to optimize a highly efficient mRNA transfection into dendritic cells (DC) derived from esophageal squamous cell carcinoma (ESCC) patients. Applying an electroporation technique, in vitro synthesized Green Fluorescent Protein (GFP) mRNA was transfected as an indicator into the DCs derived from a healthy donor. Flow cytometry revealed 84.9% transfection efficiency for DC...

متن کامل

Statistical Modeling for Oblique Collision of Nano and Micro Droplets in Plasma Spray Processes

  Spreading and coating of nano and micro droplets on solid surfaces is important in a wide variety of applications including plasma spray coating, ink jet printing, DNA synthesis and etc. In spraying processes, most of droplets collide obliquely to the surface. The purpose of this article is to study the distribution of nano and micro droplets spreading when droplets impact at an oblique a...

متن کامل

Transfection of 6-23 A Rat Thyroid Carcinoma Cell Line,with the Neuropeptide YcDNA

Neuropeptide Y (NPY) is a 36 amino acid peptide found throughout the central and peripheral nervoussystem of rat and human. NPY has been proposed to play an important role in satiety. The aim of this studywas to produce cell lines that secrete high levels of bioactive NPY. For this purpose, the complementaryDNA (cDNA) that encodes NPY was isolated by PCR. The cDNA was then clo...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

عنوان ژورنال:

دوره 2  شماره 

صفحات  -

تاریخ انتشار 2012