Cell Electroporation upgrade Model LE+

Catalog No.	11-0101
Product Name	Cell Electroporation upgrade Model LE+
Supplier Name	CELETRIX LLC
Brand Name	Celetrix
Summary	T Cell electroporation upgrade model LE+ (11-0101) is new cell line mode and PBMC mode. can be used to transfect a variety of mammalian cell line, especially PBMC, with high-efficiency and low-toxicity electroporation. The T cell electroporation system can maintain a high level of T cell survival, allowing immunotherapy applications such as CAR and TCR T-cell generation and CRISPR knockdown of T-cell genes. Keywords: cell electroporation, CHO electroporation transfection, CHO stable cell line development, NK cell electroporation machine, T cell electroporation machine, recombinant stable cell line development.
Shipping	The T Cell electroporation upgrade model LE+ (11-0101) is shipped at ambient temperature.
Stability & Storage	The T Cell electroporation upgrade model LE+ (11-0101) is stable at room temperature.
Note	Products will be shipped from the warehouse in USA. Promotion is running from time to time. Welcome to send a request for quote to message@sydlabs.com.
Order Offline	Syd Labs, Inc. 4 Avenue E, Hopkinton, MA 01748 USA. Phone: 1-617-401-8149 Fax: 1-617-606-5022 Email: message@sydlabs.com.

Description

11-0101: T Cell electroporation upgrade model: LE+

1、New cell line mode and PBMC mode;
2、Voltage range 300 – 1500 V;
3、optimized for large plasmid electroporation;
4、For 20ul, 100ul and 200ul electroporation tubes

Simplified operation with single adjustable parameter
For mammalian cells, with DNA, RNA, siRNA or proteins.
Integrated oscilloscope, electrical current pulse display.

Our T cell electroporation system is the only one that maintains high level of cell survival and expansion, allowing immunotherapy applications such as CAR-T, TCR-T generation and CRISPR knock-down of T cell genes. It is hard to transfect human T cells with other methods, especially for plasmids. Using our electroporation models LE+ and EX+, T cells can be transfected any time from the fresh PBMC stage to the stimulated or cultured stage. The T cell electroporation efficiency using our electroporation machines is the highest compared to any other electroporation methods on the market.

For example, using our electroporation machines, it was shown that the transfection efficiency to human PBMC with a GFP plasmid after 24 hours was over 80%; it was also demonstrated that after human PBMC transfection of CAR-T2A-GFP with the SB transposon, the human PBMC CAR expression was 65.9% after 14 days while cells form clusters and proliferate similar to the un-electroporated cells.

Viral vectors have been the predominant delivery vehicles in immunotherapy, since the other existing electroporation methods fared poorly with immune cells. Our new high-performance electroporation technology swiftly electroporates unstimulated PBMC cells and supports fast cell expansion, enabling clinical T cell transfection for CAR-T therapy.

Genome editing with Cell Electroporation Model LE+ and Model LE+
With Celetrix electroporation, we can deliver RNP (ribonucleoprotein complex containing Cas9 protein and gRNA) or Cas9/gRNA plasmid into cells to perform gene editing.

RNP electroporation has very low cell toxicity and the efficiency of gene editing can usually reach over 90%, with no apparent cell type limitation. For primary T cells, NK cells and stem cells, Cas9 plasmids have a high toxicity upon electroporation and we recommend using RNP on these cells.

T cell gene editing can be done with either unstimulated T cells (PBMC) or stimulated T cells, using different voltages. On the LE+/EX+ model machines, we can use the PBMC mode to electroporate RNP into PBMCs with a high voltage. Stimulated T cells are electroporated at 2-3 days of stimulation time and the voltage is similar to common cell lines, much lower than the voltage of PBMCs. For example, we can knock out the TCR using 2-3 day stimulated T cells and the efficiency of knockout is 92% and cell growth is not impaired.

T cell, NK cell and CD34+ HSC cells are important for gene and cell therapy applications. Our technical support can make gene editing in these cells very straightforward for you. Also, we have a large scale single tube electroporation machine that can process a 10 ml sample, or 2x10E9 cells in a single zap.

iPSC cells are also similar to common cell lines with Celetrix electroporation. RNP electroporation can achieve high efficiency gene editing in iPSCs, as shown with AAVS1 and OCT4 genes. One easy way to achieve high gene editing efficiency is to use relatively high concentration of RNP, such as 10 ug Cas9 and 5 ug gRNA in a 20 ul electroporation.

Plasmid based gene editing is suitable for cell lines. The cell toxicity of large plasmids containing Cas9 gene and gRNA gene can be tolerated by cell lines. The gene editing efficiency from plasmid electroporation is usually lower than the efficiency from RNP electroporation. However, we can use drug resistant gene selection to improve the gene editing efficiency. For example, THP-1 cells was electroporated with a plasmid containing Cas9, gRNA, GFP and puromycin-resistance genes and cells were all positive in GFP after puromycin selection, with a high rate of gene editing.

Normally it takes long time to do gene editing with plasmids. We have developed protocols to perform quick gene editing, even with plasmids. Because of the high cell viability and efficiency of Celetrix electroporation, drug selection can be done for a short period at the transient transfection stage and the remaining cells carry gene editing without stable integration of the plasmid. This new method speeds up plasmid based gene editing and allows sequential editing of multiple genes.

T Cell Immunotherapy with Celetrix electroporation
Human T cells have been hard to transfect with other methods, especially for plasmids.

With Celetrix electroporation, T cells can be transfected any time from the fresh PBMC stage to the stimulated or cultured stage. The Celetrix efficiency is highest compared to all other electroporation methods on the market. Our technology is also the only one that maintains high level of cell survival and expansion, allowing immunotherapy applications such as CAR, TCR-T generation and CRISPR knock-down of T cell genes.

Human PBMC transfection with GFP plasmid

Transfection efficiency with GFP plasmid after 24 hours is over 80%

Human PBMC transfection of CAR-T2A-GFP with SB transposon

4 days

14 days

20 days

14 days

Human PBMC CAR expression at 65.9% after 14 days.
Cells form clusters and proliferate similar to un-electroporated cells.

Electroporation—– The Future of CAR-T Therapy

Viral vectors have been the predominant delivery vehicles in immunotherapy, since the other existing electroporation methods fared poorly with immune cells.
The new high-performance Celetrix technology swiftly electroporates unstimulated PBMC cells and supports fast cell expansion, enabling clinical T cell transfection for CAR-T therapy.

References:

Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection: The resulting mixture was immediately transferred to 20-μL electroporation tubes and subjected to electroporation condition (voltage = 500 V, time = 20 ms) within an electroporator (Celetrix CTX-1500A LE, USA) and then gently transferred into pre-warmed Xvivo 15 medium without antibiotics.

Efficient generation of locus-specific human CAR-T cells with CRISPR/cCas12a: Electroporate using a Celetrix CTX-1500A LE electroporator. For primary T cells, electroporation is conducted at 420 V for 20 ms at one pulse.

Abstract 4064: Optimization of the transposon expression system for electroporation based gene delivery: However, in the actual application of the transposon systems such as the piggyBac (PB) system and the Sleeping Beauty (SB) system, it is quite challenging to achieve a high delivery efficiency and a high cell viability. We found that in addition to the choice of electroporation methodology, vector design and cell culture are two most important aspects that need to be optimized. Vector design dictates protein expression level as commonly anticipated, it also dictates the expression timing. We found that two distinct designs give two very different expression timing, one with initial high efficiency of expression which tapers down to about half, and another one with very low initial expression level and it goes up to a very high level after two weeks. We propose that the differential expression profiles of the vector systems can be applied to different circumstances to achieve better therapeutic effects.

Celetrix electroporation machines:

Cell electroporation basic model SP100, catalog No. 11-0103, simplified single pulse for all cell types; for 20 ul, and 100 ul electroporation tubes

Cell electroporation upgrade model LE+, catalog No. 11-0101, new cell line mode and PBMC mode. for 20 ul, 100 ul, 120 ul, and 200 ul electroporation tubes.

Cell electroporation upgrade model EX+, catalog No. 11-0106, new cell line mode and PBMC mode. for 20 ul, 100 ul, 120 ul, 200 ul, 600 ul, and 1 ml electroporation tubes.

Cell electroporation large-scale model SLT, catalog No. 11-0104, for 200 ul, 1 ml, 5 ml, and 10 ml electroporation tubes.

Cell electroporation upgrade model EX+ with the electrofusion function, catalog No. 11-0106F, for 20 ul, 100 ul, 120 ul, 200 ul, 400 ul, 600 ul, and 1 ml electroporation tubes. Model EX+ and Model SLT can be equipped with optional fusion accessories for hybridoma generation. Mouse spleen cells can be fused with myeloma cells such as SP2/0 and P3X63Ag8 with high efficiency.

UHV Transformer (Ultra High Voltage), catalog No. 11-0201, bacteria cells: 20 ul, 100 ul, and 600 ul electroporation tubes; yeast cells: 20 ul, 100 ul, 200 ul, and 1000 ul electroporation tubes.

UHV-Plus (Ultra High Voltage), catalog No. 11-0202, bacteria cells: 20 ul, 100 ul, and 600 ul electroporation tubes; yeast cells: 20 ul, 100 ul, 200 ul, and 1000 ul electroporation tubes; mammalian cells: 20 ul and 200 ul electroporation tubes.

Please remember our product information: Cell electroporation upgrade model LE+ catalog number: 11-0101 Celetrix.

Additional information

Ship from Country	USA
Size	1 Set

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