Magnetofection

Description

Magnetofection™ is a novel, simple and highly efficient method to transfect cells in culture. It exploits magnetic force exerted upon gene vectors associated with magnetic particles to draw the vectors towards, possibly even into, the target cells. In this manner, the full vector dose applied gets concentrated on the cells within a few minutes so that 100% of the cells get in contact with a significant vector dose. This has several important consequences:

  • Greatly improved transfection rates in terms of percentage of cells transfected compared to standard transfection.
  • Up to several thousand fold increased levels of transgene expression compared to standard transfections upon short-term incubation.
  • High transfection rates and transgene expression levels are achievable with extremely low vector doses, which allows to save expensive transfection reagents.
  • Extremely short process time. A few minutes of incubation of cells with gene vectors are sufficient to generate high transfection efficiency, compared to several hours with standard procedures.

Technology

Magnetofection™ Reagents

As the manufacturer of the Magnetofection™ technology, chemicell offers two types of ready-to-use Magnetofection™ reagents.

PolyMAG is a universally applicable magnetic particle preparation for high efficiency nucleic acid delivery. It is mixed in a one-step procedure with the nucleic acid to be transfected and has been used successfully with plasmid DNA, antisense oligonucleotides and siRNAs.

CombiMAG is a magnetic particle preparation designed to be combined with any commercially available transfection reagent such as polycations and lipids and can be associated with plasmid DNA, antisense oligonucleotides, siRNAs or viruses. It allows you to create your own magnetic gene vector based on your favourite transfection reagent.

Purchaser Notification
The Magnetofection™ reagents and all of its components are developed, designed, intended and sold for research use only. They are not to be used for human diagnostic or any drug intended.
Magnetofection™ are registered trademark

Nucleic Acids Dose Response and Transfection Kinetics

Transfection kinetics: NIH 3T3 cells were incubated with GenePorter™ (Gene Therapy Systems) ± CombiMAG with and without positioning on the MagnetoFACTOR plate for the indicated time spans. Luciferase expression was assayed after 24 hours.

Dose response profile in NIH 3T3 cells using Lipofectamine™ (Invitrogen) ± CombiMAG with and without positioning on the MagnetoFACTOR plate for 15 min. Luciferase expression was assayed after 24 hours.

Applications

Magnetofection™ is generally applicable for adherent cells and has been tested with a variety of immortalized cell lines and primary cells listed below. If a particular cell type or cell line is not listed this does not mean that Magnetofection™ would not work. Also for the cells listed, some reagents have not been tested so far, as indicated by “n.d.” (not determined).

The CombiMAG reagent can be combined with any polycationic and lipidic transfection reagent, and also with adenoviral and retroviral vectors. In some cases, references are made in the footnotes to very successful combinations with commercially available reagents that have been tested so far.

Type of nucleic acid / virus PolyMAG CombiMAG
Plasmid DNA yes yes
Antisense Oligonucleotides yes yes
siRNA yes yes
Adenovirus n.d. yes
Retrovirus n.d. yes

Effectene (Qiagen)

Effectene + CombiMAG

Primary human umbilical vein endothelial cells positioned on the MagnetoFACTOR plate were incubated for 15 min with a Cy3 fluorescencelabeled antisense-oligonucleotide complexed with Effectene™ (Qiagen; left) or Effectene™ + CombiMAG (right).

(Data kindly provided by F. Kroetz. Ludwig-Maximilians University Munich)

Adherent cells
Cell Line Cell Type Source PolyMAG CombiMAG* Reference Number
HeLa cervix carcinoma human + + 9.0309.0249.0729.0129.1119.0039.0059.016
HEK 293 kidney human + + 9.0139.0649.054
BHK kidney hamster n.d. +
CHO ovarian hamster + + 9.0369.0029.0059.006
NIH 3T3 fibroblasts mouse + + 9.0039.0729.0919.0029.0059.004
16HB140 airway epithelium human + n.d. 9.010
EJ28 bladder cancer human + n.d.
HBL-100 breast human n.d. +
MCF7 breast adenocarcinoma human + + 9.0149.0409.0419.115
MCS7 breast adenocarcinoma human + n.d.
HCT 116 colon adenocarcinoma human n.d. +
U373 astrocytoma human + n.d. 9.056
U87 glioblastoma human + n.d. 9.056
U251 glioblastoma human + n.d. 9.056
T98G glioblastoma human + n.d. 9.056
YK6-1 glioblastoma human + n.d. 9.056
YH-13 glioblastoma human + n.d. 9.056
A549 alveolar basal epithelial human + + 9.0759.101
A172 glioblastoma human + n.d. 9.0569.074
LN229 glioblastoma human + + 9.074
LN18 glioblastoma human + + 9.074
SW480 colon adenocarcinoma human n.d. +
LoVo colon adenocarcinoma human n.d. +
HCT15 colon adenocarcinoma human n.d. +
CEMx174 hybrid T/B-lymphocytes human n.d. + 9.060
CHO ovarian human + +
A431 epidermoid carcinoma human n.d. +
HT-1080 fibrosarcoma human + n.d.
HFF foreskin fibroblast human + +
Hep G2 hepatocellular carcinoma human + + 9.0129.048
HaCat keratinocytes human n.d. +
293 T kidney human + + 9.030
293 T-17 kidney human n.d. + 9.030
SK-MES-1 lung carcinoma human + n.d.
MRC 5 lung, embryonic human + +
MeWo melanoma human n.d. +
NYGM cerebral glioblastoma human n.d. + 9.056
SK-MEL-28 melanoma human n.d. +
MEP mammalian epithelial human n.d. + 9.047
MOLT-4 acute lymphoblastic leukemia human n.d. + 9.044
SHSY-5Y neuroblastoma human n.d. + 9.1119.050
SV40 transformed fibroblasts human + n.d. 9.027
A549 Non-small cell lung carcinoma human n.d. + 9.0579.101
SaOS-2 osteo sarcoma human n.d. +
BTK-143 osteo sarcoma human n.d. +
BEAS-2B lung epithelial cells human + n.d. 9.012
181RDB pancreatic human n.d. +
HN12 Head, neck carcinoma human n.d. + 9.028
PC-3 prostate carcinoma human n.d. +
H9 lymphoblastoid human n.d. + 9.077
H295R adrenocortical carcinoma human n.d. + 9.0589.0599.037
HOS osteosarcoma human n.d. + 9.077
HSG salivary gland, submandibular human n.d. +
He99 lung cancer human + + 9.100
HMEC-1 microvascular endothelial human + n.d. 9.0389.092
NCI-H441 lung adenocarcinoma epithelial human + + 9.073
NCI-H292 mucoepidermoid carcinoma human + n.d. 9.0659.0959.112
NCI-H82 small cell lung cancer human + n.d.
SK-N-BE2 neuroblastoma human + n.d. 9.031
MDAMB231 breast adenocarcinoma human + n.d. 9.035
ECV-304 urinary bladder carcinoma human n.d. +
11.5dpcXY gonad explants mouse n.d. + 9.109
Cal27 oral adenosquamous carcinoma mouse n.d. + 9.028
CT-26 colon carcinoma mouse + n.d. 9.0039.005
C127 murine mammary tumor mouse n.d. + 9.042
F9 embryonal carcinoma mouse + n.d.
L929 fibroblast, connective tissue mouse + n.d. 9.016
MEF fibroblast mouse + n.d. 9.0399.098
HT-22 hippocampal neuroblast mouse n.d. +
mIEnd Mesenteric lymph node mouse n.d. + 9.085
mIC-(cl2) intestine mouse n.d. +
B16F10 melanoma mouse + n.d. 9.0769.005
N2a neuroblastoma mouse + n.d. 9.0639.086
NS20Y neuroblastoma mouse n.d. +
bTC-tet B-cell line mouse + + 9.020
Hepa1c1c7 murine hepatoma mouse + n.d. 9.084
SVEC murine cell line mouse n.d. + 9.028
OP9 bone marrow-derived stromal mouse n.d. + 9.047
SM 10 trophoblast mouse n.d. +
STC-1 enteroendocrine I cells mouse + + 9.033
A7r5 aortic smooth muscle rat n.d. + 9.094
PC12 adrenal pheochromocytoma rat + n.d.
PC6-3 daughter cell line of PC12 rat + n.d.
C6 glioma rat n.d. +
H4IIE hepatoma rat n.d. + 9.062
L6 skeletal muscle cell rat + n.d.
RIE1-α5 intestinal rat + n.d. 9.088
HNSCC Head, neck carcinoma rat n.d. + 9.0159.0289.051
RV vein rat + n.d.
COS-7 kidney monkey + + 9.030
CV-1 kidney monkey + n.d.
COS-1 kidney monkey n.d. +
Vero kidney monkey n.d. + 9.0459.0469.016
B11 (UV) resistant ovary hamster n.d. + 9.028
V79 lung fibroblasts hamster + n.d.
PT-11 kidney bovine n.d. +
MDCK kidney canine + n.d. 9.067
CRFK kidney cat n.d. +
VSa13 bone (chondrocyte-like) fish + n.d.
AM-C6SC8 kidney pig n.d. +
fibroblasts xenopus + n.d.
EMC mesenthelial. Cells (epicard) n.d. +
Suspension cells
Cell Line Cell Type Source PolyMAG CombiMAG* Reference number
THP-1 acute myeloid leukemia human n.d. +
Jurkat acute T-cell lymphoma human n.d. + 9.0449.073
BL-41 B-cell lymphoma human + n.d.
K562 chronic myeloid lymphoma human n.d. + 9.002
P815 mastocytoma mouse n.d. +
U937 histiocytic lymphoma human + +
Primary cells
Cell Type Source PolyMAG CombiMAG* Reference Number
endothelial cells (cord blood) human + n.d. 9.054
aortic endothelial cells human + n.d. 9.0289.032
epithelial (HUVEC) human + n.d. 9.0299.0349.0929.1139.1149.0039.0079.0089.0219.0299.0349.0059.004
fibroblasts human + +
fibroblasts, diploid human + +
gastric gland human + +
adherent gastric cells human n.d. + 9.069
gastric myofibroblasts human + n.d. 9.043
glioma human + n.d.
keratinocytes human n.d. + 9.005
LNCaP human n.d. +
lymphocytes human + + 9.0059.102
mammal epithelium human n.d. +
nasal airway epithelium human + + 9.005
pancreatic tumor human + n.d.
peripheral blood lymphocytes human + + 9.0059.102
stroma cells (endometrium) human n.d. +
T-cells human + n.d. 9.1029.089
chondrocytes human + + 9.1079.0229.024
trophoblastic cells human n.d. +
fibroblasts (MEF) mouse n.d. + 9.0189.053
neurons mouse + + 9.071
vagal afferent neurons mouse n.d. + 9.071
adherent gastric cells mouse n.d. + 9.061
peripheral blood lymphocytes mouse n.d. +
myoblasts mouse n.d. + 9.053
T-cells mouse + n.d.
hippocampal neurons rat + + 9.0179.0709.0509.0529.0939.0909.1059.104
aortic smooth muscle cells rat n.d. + 9.094
cardiomyocyte rat n.d. + 9.055
hepatocytes rat n.d. +
epithelial (HUVEC) rat + + 9.0289.054
cortical neurons rat n.d. + 9.0529.0639.097
aortic endothelial cells bovine + n.d.
carotid artery smooth muscle bovine + n.d.
chromaffine cells bovine n.d. +
lens bovine + n.d.
airway epithelium pig n.d. +
chondrocytes pig + +
fibrochondrocytes pig + +
pSM, smooth muscle pig n.d. +

* Corresponds to successfully transfected by CombiMAG in association with different commercial transfection reagents such as FuGENE™ (Roche), Lipofectamine™, METAFECTENE™ (Biontex GmbH) and DMRIE-C™ (Invitrogen); adenovirus; Effectene™ (Qiagen)

n.d. = not determined

More Via http://www.chemicell.com/products/Magnetofection/Magnetofection_separation.html