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Newsletter_2013-05

 

 

Request a NEPA21 Demonstration

 

 

What is Sonoporation SP100 Sonoporator Hybridoma Production Algae EP Nepa21 v Amaxa Nepa21 v Neon Cationic microbubbles on cells

Request a DEMONSTRATION

NEPA21 Electroporator

3-Step In Vitro and In Vivo Electroporation Technology

 

 

 

The NEPA21 system
is our latest and most sophisticated electroporation device to-date.  It
is the s
uccessor to our hugely successful CUY21EDIT and CUY21SC
electroporator range.  The NEPA21 is based on the same platform and core
technology that made the EDIT and SC devices leaders in their
class. 

The NEPA21:

–  Since launch last year in Japan, has taken over
70% of the Japanese electroporation transfection market.

–  Unique to the NEPA21 is its 3-Step electroporation
protocol.

Step
1
: a high-voltage short duration pulse

–  Which
opens the cell membrane

Step
2
: a low-voltage longer duration pulse

–  Which
drives the pay-load (e.g., DNA) in one direction

Step
3
: reverses the polarity of the pulse and drives the pay-load in the opposite
direction

 

–  Competing devices merely store charge on a
capacitor and brutally ‘dump’ that charge in one pulse. 

o Effectively
and uniquely, the first two steps of the NEPA21 electroporation process
ameliorate the destructive impact of the delivery of an electrical pulse.

o  Together,
these first two steps account for the increased Viability statistics achievable with the NEPA21

Enhanced
Transfection Efficiency is achieved by applying the integrated and
unique Polarity
Exchange function
, Step 3. Link
to photo results

o  Voltage
Decay can also be programmed into the electroporation protocol – another
important differentiating feature of the NEPA21 compared with other
devices.  We have found this feature to contribute to both the enhanced
Viability and Transfection Efficiency obtainable with the NEPA21.

 

–  Covers the full  In Vitro
application range.

 

          o  For
both Suspended Cells and Adherent Cells

o  Compared
with other leading systems, it costs significantly less per electroporation to
run the NEPA21 system.  Less than Euro 2.00 per electroporation (if you
only use a cuvette once)

o  There
are no hidden extra costs.  The system does not require expensive
buffers.

 

 

– 

Covers the full In Vivo
application range

o  Previously,
if you wished to cover the full in vivo application range with our former
systems you had to purchase both the CUY21EDIT and CUY21SC
devices. 

For some laboratories this proved an expensive option.

o  But
now with the NEPA21 you get the

full in vivo application range in the
one device.  The NEPA21 combines in the one device BOTH the wider voltage
range
of the CUY21EDIT and the enhanced sensitivity at lower voltages
of the CUY21SC.

 

–  The NEPA21
can still be set-up to output the same single output-pulse wave-form found in
the CUY21EDIT and CUY21SC devices.

 

–  Unlike competing devices, the NEPA21 is what
we call a current controlled device.

o  Bearing
in mind the formula [V = IR]:  What this
means is that the target sample is first tested for R,
and then the delivered current is precisely controlled to obtain the
appropriate V.  The net effect of this is
that the electric fields delivered into the target are much more highly
controlled which, in turn, makes for more precise electroporation from one
experiment to the next and enhanced Viability and Transfection
Efficiencies.

o  Many
of the competing devices on the market are what we term voltage controlled
devices
. What this means is that researchers (when they are
performing their experiments) are encouraged to almost exclusively focus on
voltage settings.  Such a ‘voltage only’ approach ignores the biological
fact that current varies greatly with the impedance of the target.  The
net effect is that many competing devices simply store charge on a capacitor
and then release the energy into the target without regard for or measuring the
resistance (impedance). 

 

–  The NEPA21 delivers a precise Square Wave
throughout pulse delivery.

o  For
many competing devices, the quality of the square wave decays during the
delivery of the pulse.  This is not the
case for the NEPA21.

 
V:    Viability
TE:    Transfection Efficiency

 

NEPA21 Cell Transfection Data

 

Cell Lines: With Electroporation Cuvettes


Cell Line
DescriptionSpecies  Characteristics V TE  Photo
 HUMAN
 HeLa Human Cervical Carcinoma Cells Epithelial, Immortalized
Adherent
95% 95%  YES
 HeLa-K Human Cervical Carcinoma Cells
Epithelial, Immortalized
Adherent
90% 90%  
 293
(HEK293)
Human Embryonic Kidney Cells Epithelial , Adherent
Adenovirus type 5
90%
90%  
 293
(HEK293)
Human Embryonic Kidney Cells  Epithelial , Adherent 90%  70%
Yes
 293T
(HEK293T)
Human Embryonic Kidney Cells Epithelial , Adherent
SV40 large T antigen
90% 95%  Yes
 TIG-3 Human Embryonic Lung Fibroblasts

 

Diploid, Normal

Adherent
 90%  80%
Yes
 TIG-7 Human Embryonic Lung Fibroblasts Diploid, Normal
Adherent
 89%
(FACS)
76%
(FACS)

Yes
MRC-5 Human Embryonic Lung Fibroblasts  Diploid, Normal
Adherent
85% 90%  Yes
SUSM-1 Human Fibroblasts Fetal Liver, Epithelial, Normal, Immortalized  77%
(FACS)
 71%
(FACS)

Yes
 KMST-6 Human Fibroblasts
Whole Embryo, Epithelial

Normal, Immortalized

 

 70%  60%
Yes
 hTERT-BJ1 Human Fibroblast Normal, Immortalized, Adherent  Good   31%
(FACS)
 
 HUVEC Human Umbilical Vein Endothelial Cells  Normal, Adherent  70% 75%
 HT1080 Human Fibrosarcoma Cells
Connective Tissue

Acetabulum, Adherent

 

 93% 81%
Yes
 MIA-PaCa-2 Human Pancreatic Carcinoma cells  Epithelial , Adherent  80%  77%
Yes
 HepG2 Human Hepatoma cells Epithelial , Adherent  80%  76%
Yes
 HuH-7 Human Hepatoma Cells Epithelial , Adherent  70%  60%
Yes
 H69 Human Small-Cell Lung Cancer cells  Adherent  90%  85%  Yes

 

H1299

(NCI-H1299)
Human Lung Cancer Cells
Non-Small, Epithelial
Adherent
 90%  90%  
 HSC-2 Human Squamous Carcinoma Cells Oral , Adherent  90-95%  98%
Yes
 HSC-3 Human Squamous Carcinoma Cells Tongue , Adherent  90-95%  98%
Yes
 HSC-4 Human Squamous Carcinoma Cells Tongue, Adherent  80%  34%
Yes
 Ca9-22 Human Squamous Carcinoma Cells Gingival, Adherent  60%  60%
Yes
 HGF Human Gingival Fibroblasts Normal, Adherent  Excellent  Excellent
Yes
 MCF-7 Human Breast Cancer Cells Epithelial, Adherent  90%  90-95%
Yes
 T47D Human Breast Cancer Cells Ductal Carcinoma
Epithelial, Adherent
 90%  80-90%  
 BT-20 BT-20 Human Brest Cancer Cells  Normal, Mammary Epithelial, Adherent  70%  80%  Yes
 MCF-10A Human Breast Cells Normal, Mammary Epithelial, Adherent  Excellent  Excellent
Yes
 NUGC-3 Human Gastric Carcinoma Cells Adherent  73%
(FACS)
 68%
(FACS)
 
 A549 Human Lung Adenocarcinoma Cells Epithelial, Adherent  80-90%  90%  
 TE-1 Human Esophageal Carcinoma Cells Adherent  80-90%  41%
Yes
 TE-8 Human Esophageal Carcinoma Cells Adherent  80-90%  40%  Yes
 LNCaP Human Prostate Carcinoma Epithelial, Adherent  71%
(FACS)
 90%
(FACS)
 
 PC-3 Human Prostate Cancer Cells Epithelial, Adherent  90%   95%   
 RKO Human Colorectal Carcinoma Cells Adherent  80%  40%  
 SKOV-3 Human Ovarian Carcinoma Cells Adherent  90%  90%
Yes
 OVCAR-3 Human Ovarian Carcinoma Cells Adherent  90%  79%
Yes
 RMG-1 Human Ovarian Clear CellAdenocarcinoma Adherent  97%  67%
Yes
 CCS Clear Cell Sarcoma Adherent  47%  55%  
 SK-N-SH Human Neuroblastoma Cells Epithelial, Adherent  95%  95%
Yes
 SH-SY5Y Human Neuroblastoma Cells Adherent  60%  90%  Yes
 KG-1-C Human Oligodendroglial Cells Adherent  85%  60%
Yes
 A172 Human Glioblastoma Cells Adherent  80-90%  70%  Yes
 1321N1 Human Astrocytoma Cells Adherent  80%  80% Yes
 HaCaT Human Keratinocyte Cells Normal, Immortalized
Adherent
 40%  80%  Yes
 CCD1079sk Human Skin Fibroblasts Adherent  40%  53%
(FACS)
 Yes
 iHAM-4 Human Amniotic Mesenchymal Cells Adherent  59%  95%  
 iHAE-7 Human Amniotic Epithelial Cells Adherent  70%  40%  
 HTR-8/Svneo Human Trophoblast Cells  Adherent  95%  67%  Yes
 RPE Retinal Pigment Epithelium Cells Adherent  90%  70%  
 RPTEC Renal Proximal Tubule Epithelial Cells  Adherent  70%  85%  
 UT-7/TPO Human Megakaryocytic  Suspension  51%  46%  Yes
 Jurkat Human T-cell Leukemia Cells Round Single, Blood, Suspension  90%  85%  Yes
 Nalm-6 Human B-cell Precursor Leukemia Cells Blood, Suspension 67%
(FACS)
 70%
(FACS)
 
 MY Human Acute Lymphocytic Leukemia Cells Myelomonocytic, Suspension  94%
(FACS)
 22%
(FACS)
 
 PALL-2 Human Acute Leukemia Cells Acute Lymphoblastic  60%  27%
(FACS)
 
 KG-1
Human Acute Myeloid Leukemia  Suspension  50%  50%  Yes
 USCD/AML1 Human Leukemia Cells Suspension  50%  50%  
 Namalwa Human Burkitt’s Lymphoma Cells B lymphocyte, Blood
Suspension
 70% 75%  
 Raji Human Burkitt’s Lymphoma Cells B lymphocyte, Blood
Suspension
 74%
(FACS)
 53%
(FACS)
 
 Mutu I Human Burkitt Lymphoma Cells EBV Positive, Suspension  87%
(FACS)
 91%
(FACS)
 Yes
 Mutu III Human Burkitt Lymphoma Cells EBV Positive, Suspension  54%
(FACS)
 92%
(FACS)
 
 HL-60 Human Promyelocytic Leukemia Cells Suspension  80%  80%  
 LCL Human Burkitt’s Lymphoma Cells Immortalized, Suspension  55%
(FACS)
 43%
(FACS)
 Yes
 Ramos-
BlueTM
Human Burkitt’s Lymphoma Cells B lymphocyte, Suspension  80%  55%  Yes
 KU812 Human basophilic leukaemia Cells Suspension  96%
(FACS)
 42%
(FACS)
 
 K562 Human Chronic Myelogenous Leukemia Cells Suspension  91%
(FACS)
 99%
(FACS)
 Yes
 THP-1 Human Acute monocytic Leukemia Cells Suspension  76%  63%  Yes
MOUSE/RAT        
 NIH/3T3 Mouse Embryonic Fibroblasts Swiss NIH , Embryo
Adherent
 74%
(FACS)
 82%
(FACS)
 Yes
 3T3-L1 Mouse Preadipocyte Fibroblast like  90% 90%  Yes
 MEF Mouse Embryonic Fibroblasts Adherent  80%  90%  Yes
 MEF Mouse Embryonic Fibroblasts
 Adherent  75%  70%  Yes
 P19C6 Mouse Embryonic Carcinoma Cells  Adherent  90%  50%  Yes
 L Mouse Fibroblasts Adherent  90% 65%  Yes
 MC3T3-E1 Mouse Osteoblastic Cells Fibroblastic, Adherent  85% 75%  Yes
 LLc1

(LL/2)

 

Lewis lung Cell Carcinoma 1 Cells Adherent  87%
(FACS)
 81%
(FACS)
 Yes
 C2C12 Mouse Myoblast Cells Adherent  80%  70% Yes
 4T1 Mouse Breast Cancer Cells Adherent  90%  95%  Yes
 Colon-26 Mouse colon adenocarcinoma Cells Adherent  95%  90%  Yes
 MS-1 Mouse Pancreatic Endothelial Cells Adherent  90%  90%  
  ddy Mouse Endometrial Cells Fibroblast, Adherent  60%  80%  
 Neuro-2a Mouse Neuroblastoma Cells  Adherent  90%  90%  Yes
 BV-2 Mouse Microglial Cells  Adherent  92%  60%  Yes
 WR19L Mouse T-cell lymphoma Cells Suspension  92%
(FACS)
 60%
(FACS)
 Yes
 BA/F3 Mouse pro-B cells  Suspension  90%  90%  
 RAW264.7 Mouse Macrophage-like Cells Monocyte-like, Adherent  70%  56%  Yes
 MIN6 Mouse Pancreatic Beta Cells Suspension  57%
(FACS)
 71%
(FACS)
 Yes
  Mouse Pancreatic Islet Beta Cells    60%  40%  Yes
 MEL Mouse Erythroleukemia Cells Suspension  70%
(FACS)
 50%
(FACS)
 
 XS106 Mouse Dendritic Cells Suspension  61%  45%  Yes
 mDC Mouse Myeloid Dendritic Cells Suspension  79%
(FACS)
 72%
(FACS)
 
 MC/9 Mouse Mast Cells Suspension  76%
(FACS)
 84
(FACS)
 Yes
 BMBa Mouse Bone marrow-derived basophils Suspension  45%
(FACS)
 67%
(FACS)
 Yes
 TS Mouse Trophoblast Stem Cells Adherent  59%  47%  Yes
 ATDC5  Mouse Pre-chondrocytic Cells    62%  14%  Yes
 PC12 Rat Adrenal Pheochromocytoma Cells Adherent  90%  70%  Yes
 H9c2 Rat Ventricular Myoblasts  Skeletal Muscle-like
Adherent
 71%
(FACS)
82%
(FACS) 
 Yes
 REF Rat Embryonic Fibroblasts  Adherent  90% 99% Yes
 RDF Rat Dermal Fibroblasts Adherent 90% 40%  
 C6 Rat Glioma Cells Fibroblast-like, GFAP Positive, Adherent  80%  67%  Yes
 RSC96 Rat Schwann Cells Adherent  70%  85%  Yes
 TtT/GF Rat Anterior Pituitary Cells Folliculostellate, Adherent  65%  83% Yes
 UMR106 Rat Osteoblastic Cells Adherent  80%  70%  
 HAMSTER

 

 

 CHO Chinese Hamster Ovary Cells Epithelial, Adherent  74%  90%  Yes
 CHO-K1 Chinese Hamster Ovary Cells Epithelial, Adherent  95%  95%  Yes
 CHO-S Chinese Hamster Ovary Cells Epithelial, Suspension  Excellent  56%
(FACS)
 
 VARIOUS
         
 DT40 Chicken B Cells Suspension  38%  40%
(FACS)
 Yes
 MDCK Madrin-Darby Canine Kidney Cells Epithelial-like, Adherent  90%  95%  Yes
 BFF Bovine Fetal Fibroblasts  Adherent  78%
(FACS)
 72%
(FACS)
 Yes
 A6 Xenopus Kidney Epithelial Cells  Adherent  90%  60%  Yes
 GTS9 Ginbuna Thymus Cells Adherent  50%  30%  Yes
 Vero African Green Monkey Kidney Cells  Epithelial-like, Adherent 58%  35%  
 SEC Bovine T Cells  Suspension  80% 21%
(FACS) 
 
 
       

 

 

Stem Cells, ES Cells and iPS Cells: With Electroporation Cuvettes

Cell Line Description
Species 
Characteristics V  TE  Photo
  Human Mesenchymal Stem Cells  Primary, Adherent Cord blood derived

 

78%

(FACS)
 75%

(FACS)

 

 Yes
 TS Mouse Trophoblast Stem Cells  Adherent  59%  47%  Yes
 TT2 Mouse TT2 ES Cells  Adherent  50-60%  50-60%  Yes
  Mouse ES Cells  Adherent  80%  75%  Yes
  Rat WDA ES-like Cells  Adherent  60%  80%  
  Mouse iPS Cells  Origin: Mouse Fibroblast, Adherent  89%  47%  Yes
  Mouse Neurospheres – Neural Stem Cells Derrived from Mouse SVZ    Excellent  Excellent  Yes
  Mouse Neural Stem Cells  Primary, Adherent Good  37% Yes


Primary Cells: With Electroporation Cuvettes

 Cell Line

 

Description
Species

 

Characteristics

V 

TE

 


Photo

 

HASM Human Airway Smooth Muscle Cells Primary, Adherent 90% 80% Yes
  Human Endometrial Stromal Cells Primary Adherent 95% 90% Yes
  Human Uterine Cervical Fibroblasts  Primary, Adherent  65%  90% Yes
  Human Amniotic Mesenchymal Cells  Primary, Adherent  70%  40%  
  Human T Cells  Primary, Suspension  58%
(FACS)
 90%
(FACS)
 Yes
  Mouse Cerebral Cortex Neurons
(Embryonic day14)
 Primary, Adherent  80%  70%  Yes
  Mouse Hippocampal Neurons
(Embryonic day 14)
 Primary, Adherent  80%  60%  
  Mouse Basal Ganglia Primordium
(Embryonic day 13.5)
 Primary, Adherent  91%
(FACS)
 71%
(FACS)
 
  Mouse Neural Stem Cells  Primary, Adherent  Good  37%  Yes
  Mouse Microglia Cells  Primary, Adherent  60-70%  23%  Yes
 MEF Mouse Embryonic Fibroblasts  Primary, Adherent 90%    85%   Yes
  Mouse Embryonic Skin Fibroblasts  Primary, Adherent  90%  50%  Yes
 BMMC

(BMMCs)

 

Mouse Bone Marrow-Derived Mast Cells  Primary, Suspension  64%
(FACS)
78%
(FACS)
 Yes
  Mouse Primary Liver Cells  Primary, Adherent  70%  50%  Yes
  Mouse Stromal Cells  Primary, Adherent  90%  30%  
  Rat Cerebral Cortex Neurons (Embryonic day 16)  Primary, Adherent  64%  78%  Yes
  Rat hippocampal Neurons  Primary, Adherent  60%  80%  
  Rat Bulbar Neurons
(Embryonic day 15)
 Primary, Adherent  80%  75%  Yes
  Rat Cerebellar Granule Cells  Primary, Adherent  70%  80%  
  Rat Meningeal Fibroblasts
(Postnatal day 3)
 Primary, Adherent  90%  95%  Yes
 OEC Rat Olfactory Ensheathing Cells
(Postnatal week 3)
 Primary, Adherent  93%  46%  Yes
  Chick Embryonic Fibroplasts  Primary, Adherent  80%  90%  Yes
  Chick Embryonic Cerebellar Granule Cells (Embryonic day 11)  Primary, Adherent  86%  83%  
  *Cell clusters
Chick Embryonic Cerebellar Granule Cells (Embryonic day 13)
 *Explant
Primary, Adherent
   35%  Yes
Cell Lines in Adherent Status: With Adherent-Cell Electrodes
 
 Cell Line Description
Species
 Characteristics  V  TE  Photo
 SH-SY5Y Human Neuroblastoma Cells  Adherent  90%  50%  Yes
 C6 Rat Glioma Cells  Fibroblast-like, GFAP Positive, Adherent  57%  55%  Yes

Primary Cells in Adherent Status: With Adherent-Cell Electrodes

 

 Cell Line  Characteristics   V  TE   Photo 
Human epidermal keratinocyte Cell  Primary, Adherent  90%  30%  
Mouse Cerebral Cortex Neurons

(Embryonic day 15) (6 DIV)

 

 Primary, Adherent  Excellent  Excellent  Yes
Mouse Hippocampal Neurons
(Embryonic day 14) (4 DIV)
 Primary, Adherent  60%  50%  
Mouse Hippocampal Neurons
(Embryonic day 18) (2 DIV)
 Primary, Adherent  85%  54%  Yes
Mouse Stromal Cells
(1-month cultured)
 Primary Adherent  90%  50%  
Mouse Glial Cells
(14 DIV)
 Primary Adherent  80%  50%  Yes
 Mouse Cerebral Cortex Neurons
(Embryonic day 17) (2 DIV)
 Primary Adherent  70% 60% Yes
Rat Granulosa Cell  Primary Adherent  Excellent  41%  
Rat Neurons (E17 rat cerebral cortex) After 2 DIV (on 24-well plates)  Primary Adherent  70% 60% Yes
Rat Neurons (P7 rat hippocampus)
After 11 DIV
 Primary Adherent 100% 50% Yes
 

Plant Protoplasts

 

 Cell  Characteristics  Luciferase Activity
(CPS)
Tabacco Protoplasts    20,312

 DOWNLOADS

– Hybridoma Production with ECFG21 device

–  Chlamydomonas Reinhardtti Transformation without cell-wall removal

– NEPA21 comparison with Lipofectamine, FuGENE HD and NEON

– NEPA21 Presenation

– Measurement and Display of Delivered EP Pulse – Comparison of NEPA21 and BTX ECM830

–  Comparison of NEPA21 with Amaxa-Nucleofector, Neon and Lipofectamine 2000

– Comparison of NEPA21 with Bio-Rad 

– Resistance Ranges for NEPA21 Applications 

 

 NEPA21 Videos of example applications

           In Utero Electroporation

           Direct delivery of MIF Morpholinos into Zebrafish Embryo

          In Vivo Mouse Tumor

 

 

 

Please feel free to contact us at: sales@sonidel.com

to request a DEMONSTRATION

 


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