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Category Archives: Transfection

Lungs-Targeted Transfection Reagent: Safe and Effective Delivery of Biomolecules

August 2, 2023

Today we are happy to announce the launch of a new lungs-targeted in vivo transfection reagent. The reagent is crafted to transport small molecules, DNA, RNA, and proteins to the lungs tissue, aiming to treat lung diseases, including cystic fibrosis and lung cancer. The transfection reagent is a non-viral complex comprising nanoparticle-sized cationic liposomes that play a crucial role in facilitating the efficient delivery of cargo biomolecules into lung cells while avoiding the activation of the immune response (transfection protocol and data are now available here).

Lung cancer stands as the primary contributor to cancer-related deaths and ranks among the most frequently diagnosed malignancies worldwide. During the past 14 years Altogen Biosystems successfully engineered in vitro transfection reagents optimized for many lung cancer cell lines (A549, H460, COS7, NCI-H23, 3LL, Calu3, NCI-H1975, 4T1, Calu6, H1975, etc). Building upon this experience, the company has extended its efforts to develop an in vivo lung-targeted delivery agent. In vivo transfection, unlike in vitro transfection which occurs in a petri dish, is the process of delivering cargo molecules into tissue. When it comes to lung-targeted in vivo transfection, there are several challenges due to the organ’s natural defense mechanisms, delivery efficiency, immune response, mucosal clearance, and functionality of cargo biomolecules in vivo.

Altogen Biosystem’s Lungs In Vivo Transfection Kit was functionally validated using seven CDX xenograft models: Huh-7, NCI-H226, A549, Calu-3, H460, NCI-H1975DMS273 demonstrating that reagent surpasses the inherent deposition mechanism of inhaled aerosol droplets in the lungs, enabling targeting not only on the central airways or lungs periphery but also on specific local regions within the lungs.

About us:

Altogen Biosystems is a leading provider of life science research transfection reagents. Our mission is to provide scientists with the tools needed to make new discoveries and improve human health. Altogen Biosystems’ unwavering dedication to innovation and research, coupled with its broad range of unique products and services, has helped position the company as a leader in the field of in vivo research products.

Citations and References: Altogen® Transfection Kits

Citations and References: ALTOGEN® Transfection Reagents Nature 2008 454(7203):523-7. Innate immunity induced by composition-dependent RIG-I …Saito et al [PDF] Nature Medicine 2016 22(10):1131–1139. A Long Non-Coding RNA Defines an Epigenetic Checkpoint … Wang et al [PDF] Cell Rep 2018 22(2):523. The Foxo1-Inducible Transcriptional Repressor Zfp125 … Gustavo et al [PDF] Molecular Therapy 2018. Liposome Lipid-Based… Continue Reading

Crossing Blood-Brain Barrier and In Vivo Delivery of Negatively Charged Biomolecules into Glioblastoma Enabled by Altogen's Nanoparticle In Vivo Transfection Reagent

LAS VEGAS, NEVADA – July 19, 2021 – Altogen Biosystems, leading manufacturer of in vivo transfection products, announced today that Altogen’s nanoparticle-based in vivoTransfection reagent (catalog #5031) was demonstrated to be effective for glioblastoma-targeted co-delivery of plasmid DNA (3.7 kb) and chemically modified siRNA. This transfection reagent was used in over 30 research publications, demonstrating highly… Continue Reading

Stable Cell Line

A stable cell line is a population of cells that have been genetically modified to express a specific gene or RNA molecule of interest on a stable basis. Stable cell lines are generated by introducing the gene or RNA molecule into the cells and selecting for cells that have stably incorporated the genetic material and… Continue Reading

How do I increase transfection efficiency?

Transfection is considered as a major laboratory method to integrate protein, RNA and DNA molecules into tissues and cells. Delivery of plasmid DNA molecules containing gene inserts, messenger RNA and small interfering RNA molecules that regulate gene expression (i.e. microRNA, siRNA) into the primary cells and cancer cell lines have been extensively utilized by scientists.… Continue Reading

Nanoparticle In Vivo Transfection Reagent (Altogen)

July 27, 2021 – Altogen Biosystems® is a biotechnology manufacturing company that specializes in the development and testing of optimized cell transfection reagents and kits for both in vitro and in vivo delivery. In 2018 Altogen Biosystems launched the AltoFect transfection reagent formulated for difficult-to-transfect cells (like B-cells, T-cells), and primary cell cultures that are… Continue Reading

Quantitate in vitro anti-proliferation experiments using a metabolic assay (Alamarblue, MTT) vs measuring protein abundance (Sulforhodamine B)

The need to measure cell proliferation effects is vast, including testing the effects of growth factors, novel pharmacological agents, cytotoxicity assessment or investigating cell activation.  Cell proliferation assays utilize a stain or substrate to make a correlation between the readout and the number of remaining cells post-treatment.  However, the means of assessing cell number varies… Continue Reading

What is Chemical Transfection

Chemical transfection is a method used to deliver exogenous nucleic acids, such as DNA or RNA, into cells using chemical compounds. This method involves the use of transfection reagents, which are chemical compounds that can form complexes with nucleic acids and facilitate their uptake by cells. Chemical transfection typically involves mixing the nucleic acid of… Continue Reading

What is the best antibiotic to use for stable cell selection in mammalian cells?

There is a long list of antibiotics available to researchers when applying selective pressure in the creation of a stable cell line. The choices include zeocin, hygromycin, blasticidin, puromycin and geneticin (G418). Researchers use different antibiotics due to cost or availability in their lab. However, in reference to using antibiotics for the creation of a… Continue Reading

Is it necessary to synchronize cells?

Exponentially growing cultured cells are growing at difference cell cycle stages, termed asynchronous.  Scientists are vigilant in their techniques to ensure they are using non-confluent cells prior to their experiments; thus, synchronization is not commonly employed.  However, synchronizing your cells can help improve the results of certain cell based assays such as viral integration or… Continue Reading

What is Transient Transfection

Transient transfection is a method used to deliver exogenous nucleic acids, such as DNA or RNA, into cells on a temporary basis. This method involves the use of transfection reagents, which are chemical compounds that can form complexes with nucleic acids and facilitate their uptake by cells. Transient transfection typically involves mixing the nucleic acid… Continue Reading

Difference between technical and biological replicates

The basic definitions of technical and biological replicates are as follows: Technical replicates: a test performed on the same sample multiple times; i.e., if there are triplicate non-treated samples, a technical replicate would be testing sample #1 of the non-treated multiple times Biological replicates: a test performed on biologically distinct samples representing an identical time… Continue Reading

cGMP-grade Nanoparticle Transfection Reagent for In Vivo Delivery

Altogen Biosystems is a manufacturer of life science research products that includes in vivo transfection reagents designed for tissue-targeted delivery of biomolecules. Altogen’s Nanoparticle In Vivo Transfection Reagent was launched in 2008 and used in over 30 research publications demonstrating highly efficient delivery of cargo nucleic acids (siRNA, microRNA, mRNA, plasmid DNA, and small proteins)… Continue Reading

Even cell distribution in the 6-well plate instead of a ring of cells around the edges?

Performing cell culture experiments in round well plates can often lead to troubling spatial dispersion of the cells.  This can be attributed to culture incubators having high vibrations, non-level incubators or bad practices such as swirling the plate after adding cells.  In order to achieve even distribution of cells in the wells of culture plates,… Continue Reading

CRISPR/Cas9 Transfection Optimization

Efficient delivery of the CRISPR/Cas9 protein complex can be achieved efficiently through the use of cationic lipid-based transfection reagents. Cells may undergo electroporation to increase the probability of successful transfection. Once inside a target cell, the CRISPR/Cas9 protein complex requires no additional stimulation to start modifying the cell’s genome. Genome modification will occur within one… Continue Reading

Low cell viability due to the freezing or thawing of cancer cell lines

Freeze slow, thaw fast.  The process of freezing and thawing cancer cell lines requires extreme attention to protocol.  For freezing, a good freezing media that contains DMSO is added to the correct number of cells following the manufacturer’s instructions.  The vital part of the freezing process is to ensur that the freezing process is slow. … Continue Reading

How to determine mycoplasma contamination?

Mycoplasma contamination is a common occurrence and labs must be vigilant in monitoring and combating contamination issues.  Mycoplasma are bacteria extremely difficult to detect in cultured cells by visual inspection.  Many labs do not incorporate recurring mycoplasma testing until the lab experiences a contamination incident, at which time cleaning and testing procedures are put in… Continue Reading

Forward Transfection or Reverse Transfection?

Forward and reverse transfection protocols each have their significant uses in research.  The main protocol difference between forward and reverse transfection is whether or not the cells are plated the day before transfection (as in forward transfection) or seeded at the same time of the transfection.  Forward transfection is commonly used in situations where the… Continue Reading

AltoFect Transfection Reagent for Primary Cells and Hard-to-transfect Cell Lines

Altogen Biosystems developed AltoFect™ Transfection Reagent – a new generation transfection reagent optimized for high transfection efficiency of primary cells and difficult-to-transfect cell lines. It has been successfully tested to deliver all types of nucleic acids (plasmid DNA constructs, si/miRNA, shRNA, dsRNA, mRNA, etc), small proteins, and negatively charged small molecules. Cancer cell lines and primary cell types, a… Continue Reading

AltoFect v2.0 second generation transfection reagent

AltoFect v2.0, the second generation transfection reagent for primary cell types and hard-to-transfect cell lines. AltoFect transfection reagent exhibits up to 85% transfection efficiency in difficult-to-transfect cells such as T-cells, B-cells, and primary cell cultures. The advance has opened up the path to reliable in vitro testing of realistic responses from cells that were previously… Continue Reading

Plasmid DNA and siRNA transfection protocol optimization

There are several experimental parameters that affect transfection efficiency and associated cell viability for siRNA and plasmid DNA transfection experiments: Transfection protocol (method and conditions) Health of cultured cells Purity and concentration of pDNA and/or siRNA Controls Transfection protocol Optimized transfection protocols are available for over 100 cancer cell lines and primary cell types (see… Continue Reading

Stable transfection reagents and techniques

Stable transfection, sometimes called permanent transfection, is the integration of plasmid DNA into the chromosome of cancer cell’s DNA.  The creation of a stable cell line enables the researcher to analyze long term effects of the introduced gene.  Downstream studies incorporating the stable cell line includes overexpression of the gene insert for protein production, cell… Continue Reading

How much antibiotic required for stable cell selection?

Stable cell lines are a crucial laboratory tool that over-expresses a gene of interest in order to study gene functions, screen experimental drugs or produce therapeutic proteins (i.e. recombinant antibodies).  The cell lines will divide and continue to express the inserted transgene.  Briefly, exogenous plasmid DNA is transfected into a host cell line, which is… Continue Reading

Transfection optimization

The most vital aspect of a transfection is ensuring all conditions are optimized, including transfection reagent volume, oligo concentration, cell viability and passage number, lack of negative control activity, activity of positive control and calibrated incubator temperature, humidity and percentage of CO2.  Even if all these parameters are correctly addressed, transfection efficiency must be determined… Continue Reading

Pharmacology Abbreviations

A reference table for commonly used pharmacology terms and abbreviations. Drugs   ACD Anticonvulsant drug ACR Adverse drug reaction ASA Acetylsalicyclic acid (aspirin) CD Curative dose D, dos Dose, dosage DAW Dispense as written DAWN Drug Abuse Warning Network DIG Digitalis DSB Drug-seeking behavior IND Investigational new drug INH Isoniazid (TB drug) LD Lethal dose… Continue Reading

How to determine cell viability?

Cell viability is a calculation of the number of viable or living cells within the total number of cells.  Although there are now commercially available alternatives to the historical method, trypan blue exclusion is the tried and true method commonly used in the lab.  Briefly, trypan blue dye is added to a cell suspension and… Continue Reading

What is in vivo

In vivo refers to experiments or studies that are conducted inside a living organism or cell, usually in a laboratory animal or human subject. The term “in vivo” is Latin for “in the living.” In vivo studies can be conducted on a wide range of biological and medical systems, such as diseases, drug development, or… Continue Reading

What is the benefit of using an electroporation buffer

The process of electroporation exposes the cells to a high-voltage pulse of electricity to disrupt the phospholipid bilayer of the cell membrane causing the formation of temporary pores.  Any charged molecules (e.g. RNA, DNA) are forced into the cells thru the pores. Electroporation buffers are formulations that mimic cellular cytoplasm composition; thus, enhancing pore resealing… Continue Reading

Changing transfection plate format from a 6-well to a 96-well format

Moving a transfection reaction from a 96-well plate to a 6-well plate, or vice versa, and wanting to achieve identical results is not a trivial pursuit.  It is well known that lipoplexes adhere to the plastic surfaces of cell culture wells.  As one can conclude, this decreases the amount of complexed material freely available to… Continue Reading

Importance of cell passage number and cell confluency for efficient transfection

The viability, confluency and passage number are all vital parameters for a successful transfection.  Here are helpful hints for each of these aspects: Viability Viability is the percentage of living cells in a suspension Determined using trypan blue exclusion Cells are considered to be healthy if viability is greater than 90% Confluency Confluency is the… Continue Reading

Tissue culture using flasks vs dishes for cell lines

Flasks and dishes that are cell culture coated are both acceptable consumables to use for cell culture.  Employing sterile techniques allows an experienced scientist to use either item, with pros and cons listed here for each system. Filtered Flasks Narrow opening translates to a lesser chance of contaminant entering the flask Ideal for long term… Continue Reading

Cell passage: How to correctly dilute and split cultured cells

There are many different cell culture techniques. Utilizing correct cell passaging methods is important to keep cell line in exponential growth curve, therefore making it a good model as a host for transfection experiments. “Cell passage” is a term used by other scientists to demonstrate the following process: Wash cells with PBS Detach cells from… Continue Reading

How to ship cells as a live culture?

Shipping cells to collaborators can be accomplished by either sending them a frozen aliquot on dry ice or by sending a live culture.  Sending a live culture has the advantages of the cells continuing their exponential growth to aid in timing of experiments.  This is accomplished by completely filling the flask with culture medium, closing… Continue Reading

What transfection controls do I need to include in my experiment?

Scientists understand the importance of controls in experiment. There are at least three transfection controls that should be included on every transfection plate: a positive control, negative control and non-treated control. Positive Controls Transfecting a positive control ensures that the system being utilized is working and the delivery conditions are optimal.  The results of the… Continue Reading

What is the best method to detach cultured cells?

Cultured adherent cells routinely need to be detached and collected for counting or passaging.  Detaching cells can be accomplished by either mechanical or enzymatic methods. Mechanical: cell scraping is a good option for cells that are sensitive to trypsin but can cause damage to cells; also can be used when collecting cellular components for western… Continue Reading