Cell Culture

<h2 data-section-id="xs82dy" data-start="121" data-end="167">What are additional immortalization agents?</h2>
&nbsp;
Additional immortalization agents cover recombinant lentiviral constructs delivering genes other than TERT or SV40 large T antigen used to bypass replicative senescence in primary cells, most notably HPV-16 E6/E7 (which inactivate p53 and Rb similarly to SV40T but are derived from human papillomavirus), Bmi1 (a polycomb group protein that represses the p16/CDKN2A senescence pathway), cMyc, CDK4, and HOXA/HOXB family genes. Related entities include p16/CDKN2A pathway repression (Bmi1), keratinocyte and epithelial cell immortalization (HPV E6/E7), polycomb group signaling, and combination immortalization strategies.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" role="presentation">Lenti-HPV-16 E6/E7 degrades Rb and p53 for enhanced cell immortalization.</li>
<li dir="ltr" role="presentation">Lenti-Myc T58A &amp; Ras V12 enhance transformation and proliferation assays with high-titer mutant oncogenes&nbsp;</li>
<li dir="ltr" role="presentation">Lenti-p53 &amp; Rb siRNA Virus extends cellular longevity and targets the knockdown of tumor suppressors</li>
<li dir="ltr" role="presentation">Lenti-HOX Series (RFP Tagged) for lineage-specific expansion and includes high-titer vectors like HOXB8, HOXA9, and HOXA10 vectors.</li>
<li dir="ltr" role="presentation">EBV Virus (Wild-Type) for immortalization workflows of B-lymphocytes&nbsp;</li>
<li dir="ltr" role="presentation">Lenti-CDK4 &amp; Bmi1 Virus helps maintain proliferative capacity using cell mediators</li>
</ul>
&nbsp;
<h2 data-section-id="1acs4rs" data-start="1029" data-end="1087">How to choose additional immortalization agent products</h2>
&nbsp;
HPV E6/E7 for epithelial and keratinocyte immortalization
HPV-16 E6 and E7 proteins inactivate p53 and Rb respectively, similar in net effect to SV40 large T antigen but derived from a different viral source; E6/E7-based immortalization is particularly well-documented for keratinocytes and other epithelial cell types, and is sometimes preferred over SV40T for these lineages based on published precedent.
Bmi1 for stem and progenitor cell types
Bmi1 represses the p16/CDKN2A senescence pathway through its role in the polycomb repressive complex, and has been used to extend the proliferative capacity of hematopoietic and neural stem/progenitor cells in research settings; confirm the specific cell type has documented precedent for Bmi1-based extension before relying on it as a primary immortalization strategy.
cMyc and CDK4 as proliferation-driving factors
cMyc and CDK4 overexpression can drive proliferation and contribute to immortalization, often used in combination rather than as standalone immortalization agents; review the specific combination strategy in the source literature before ordering individual components.
E6/E7 separate vs. fused constructs
HPV E6/E7 may be delivered as separate genes or as a fused open reading frame; confirm which configuration a published protocol specifies, since this can affect relative expression levels of the two proteins.
&nbsp;
<h2 data-section-id="1fljvas" data-start="3287" data-end="3312">Specifications context</h2>
&nbsp;
Additional immortalization agent lentiviruses are typically supplied at titers around 1x10^8 TU/mL, shipped on dry ice and stored at -80&deg;C, with antibiotic or fluorescent-antibiotic fusion selection markers consistent with other immortalization lentivirus products. Pack sizes for academic labs commonly consist of small-volume aliquots sufficient for multiple transduction attempts, with constructs validated by sequencing and tested free of common contaminants.&nbsp;
This sub-category sits within MBP's cell immortalization catalog alongside TERT overexpression viruses and SV40T antigen overexpression viruses. Labs constructing custom combination vectors can also review MBP's cloning reagents. MBP's specialist team can help confirm which immortalization agent has documented precedent for a specific cell type before order placement.
<a href="https://mbpinc.net/contact-us">Contact the expert team </a>at MBP today to book high-quality immortalization vectors for your lab.

Additional Immortalization Agents

MSDS

Lentiviral Vector Amplification - Protocol

Lentivector Packaging - Protocol

Lentivirus Infection - Protocol

Cell Immortalization - Handbook

Cell Immortalization Reagents - FAQ

Lenti-p53 siRNA Virus, High Titer

Lenti-p53 siRNA Virus, High Titer (LV619) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing p53 siRNA for stable knockdown of the p53 tumor suppressor, supplied as 2 × 100 µl for cell immortalization and p53/TP53 pathway research. Shipped on dry ice; store at –80°C.

Unit Quantity

Application

Expression System Type

Vector Map

Note

Caution

Shipping Condition

Lenti-Ras V12 Virus, High Titer

Lenti-Ras V12 Virus, High Titer (LV620) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing constitutively active Ras V12 (G12V), supplied as 2 × 100 µl for stable immortalization of neural stem cells, primary kidney cells, erythroblasts, and epithelial cells. Shipped on dry ice; store at –80°C.

Lenti-Rb siRNA Virus, High Titer

Lenti-Rb siRNA Virus, High Titer (LV621) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing Rb siRNA for stable knockdown of the retinoblastoma protein (pRb), supplied as 2 × 100 µl for cell immortalization and pRb/cell cycle research. Shipped on dry ice; store at –80°C.

<h2 data-section-id="6b0hnw" data-start="102" data-end="151">What are SV40T antigen overexpression viruses?</h2>
&nbsp;
SV40T antigen overexpression viruses are recombinant lentiviral particles carrying the simian virus 40 (SV40) large T antigen gene, a hexamer protein that inactivates the p53 and retinoblastoma (Rb) tumor suppressor pathways, bypassing the senescence checkpoints that normally limit primary cell proliferation. Some constructs deliver the large T antigen alone, while others include both large and small T antigens. Related entities include p53/Rb pathway inactivation, broad cross-species immortalization (human, mouse, rat, bovine, porcine, canine), karyotype/tumorigenicity characterization (soft agar assay), and antibiotic/fluorescent selection markers.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" role="presentation">Lenti-SV40 (tsA58 Mutant) for cell proliferation with inducible control and temperature-sensitive expression</li>
<li dir="ltr" role="presentation">Lenti-SV40T (Puro/Neo/No Marker) provides stable integration with versatile selection markers of highly concentrated lentiviral stocks&nbsp;</li>
<li dir="ltr" role="presentation">Adeno-SV40 Adenovirus for transient T-antigen expression with high-efficiency adenoviral vectors</li>
<li dir="ltr" role="presentation">Retro-SV40 &amp; Retro-SV40LT for special integration in proliferating cells.</li>
<li dir="ltr" role="presentation">Lenti-SV40Tt Lentivirus: Small T and Large T antigens for improved transformation.</li>
</ul>
&nbsp;
<h2 data-section-id="ykynp3" data-start="1032" data-end="1092">How to choose SV40T antigen overexpression virus products</h2>
&nbsp;
Large T antigen alone vs. large and small T antigen
Constructs delivering large T antigen alone target p53/Rb pathway inactivation specifically, while large-and-small-T constructs include additional SV40 small T antigen activity affecting protein phosphatase 2A (PP2A) signaling; confirm which form a published protocol specifies, since the two can produce different immortalization efficiencies and phenotypes.
Species applicability
SV40 large T antigen has documented immortalization success across human, mouse, rat, bovine, porcine, and canine primary cells, making it a broadly applicable option when hTERT (human-specific) is not suitable for the target species.
Selection marker and promoter
As with TERT lentivirus, SV40T constructs are available under CMV or EF1-alpha promoters with antibiotic resistance (puromycin, hygromycin, blasticidin, neomycin/G418, zeocin) or fluorescent-antibiotic fusion markers (GFP-Bsd, RFP-Puro); select based on the target cell line's existing markers and expected culture duration.
Post-immortalization characterization
Because SV40 large T antigen inactivates tumor suppressor pathways, immortalized lines should be characterized for karyotype changes and tested by assays such as soft agar colony formation and tumorigenicity testing if downstream applications require confirmation that the line has not undergone malignant transformation.
&nbsp;
<h2 data-section-id="1fljvas" data-start="3482" data-end="3507">Specifications context</h2>
&nbsp;
SV40T antigen lentivirus is typically supplied at titers around 1x10^8 TU/mL, shipped on dry ice and stored at -80&deg;C, with constructs validated by full-length sequencing and confirmed free of common contaminants. Immortalized lines derived from SV40T transduction are commonly characterized by Western blot or immunofluorescence confirming SV40-LT protein expression, alongside marker expression comparison to the original primary cells. As of 2026, SV40 large T antigen remains the most broadly cross-species-applicable immortalization approach, often selected when hTERT (human-specific) is not an option for the target species or when faster immortalization is prioritized over preserving the original phenotype.
This sub-category sits within MBP's cell immortalization catalog alongside TERT overexpression viruses and additional immortalization agents. Labs constructing custom SV40T expression vectors can also review MBP's cloning reagents. MBP's specialist team can help confirm species applicability and vector format for a specific primary cell type before order placement.
<a href="https://mbpinc.net/contact-us">Contact the expert team </a>at MBP today to get high-quality SV40T antigen overexpression viruses.

SV40T Antigen Overexpression Viruses

Lenti-SV40 Lentivirus, High Titer

Lenti-SV40 Lentivirus, High Titer (LV612) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing the complete SV40 viral genome, supplied as 2 × 100 µl for cell immortalization, SV40 biology research, viral transformation studies, and host-virus interaction research. Shipped on dry ice; store at –80°C.

Lenti-SV40 (tsA58 temperature sensitive mutant) Lentivirus, High Titer

Lenti-SV40 (tsA58 Temperature-Sensitive Mutant) Lentivirus, High Titer (LV630) by Applied Biological Materials is a 10⁹ IU/ml lentivirus expressing temperature-sensitive SV40T (tsA58) with puromycin selection, supplied as 2 × 100 µl. Active at 33°C (proliferation); inactivated at 37–39.5°C (differentiation). For conditional immortalization and stem/progenitor cell research. Shipped on dry ice; store at –80°C.

Lenti-SV40 (tsA58 temperature sensitive mutant) (No Selection Marker) Lentivirus, High Titer

Lenti-SV40 (tsA58) (No Selection Marker) Lentivirus, High Titer (LV680) by Applied Biological Materials is a 10⁹ IU/ml lentivirus expressing temperature-sensitive SV40T (tsA58) without a selection marker, supplied as 2 × 100 µl. Active at 33°C (proliferation); inactivated at 37–39.5°C (differentiation). No-selection counterpart to LV630. Shipped on dry ice; store at –80°C.

Lenti-SV40T  (No Selection Marker) Lentivirus, High Titer

Lenti-SV40T (No Selection Marker) Lentivirus, High Titer (LV665) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing SV40 large T antigen without a selection marker, supplied as 2 × 100 µl for primary cell immortalization, co-transduction, and custom selection workflows. Shipped on dry ice; store at –80°C.

Lenti-SV40T (Neo) Lentivirus, High Titer

Lenti-SV40T (Neo) Lentivirus, High Titer (LV660) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing SV40 large T antigen with neomycin (G418) resistance, supplied as 2 × 100 µl for stable primary cell immortalization with antibiotic selection. Shipped on dry ice; store at –80°C.

Lenti-SV40T (Puro) Lentivirus, High Titer

Lenti-SV40T (Puro) Lentivirus, High Titer (LV613) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus expressing SV40 large T antigen with puromycin resistance, supplied as 2 × 100 µl for stable primary cell immortalization with puromycin selection. Shipped on dry ice; store at –80°C.

Lenti-SV40Tt Lentivirus, High Titer

Lenti-SV40Tt Lentivirus, High Titer (LV614) by Applied Biological Materials is a 10⁹ IU/ml recombinant lentivirus co-expressing SV40 large T and small t antigens (SV40Tt), supplied as 2 × 100 µl for stable immortalization of challenging primary cell types. Shipped on dry ice; store at –80°C.

<h2>What are L-glutamine and amino acid supplements?</h2>
&nbsp;
L-glutamine and amino acid supplements are cell culture additives used to support cellular metabolism, growth, viability, and biosynthesis. They provide nutrients that may become depleted during culture or are required in greater quantities than those supplied by basal media alone. These supplements are commonly added to media such as DMEM, RPMI 1640, MEM, and DMEM/F-12 to optimize cell growth and maintain healthy cultures.
L-glutamine is one of the most important supplements in mammalian cell culture because it serves as a major energy source and is required for protein synthesis, nucleotide production, and glutathione synthesis. However, because free L-glutamine is unstable in solution and gradually breaks down into ammonia and pyroglutamate, many laboratories use stabilized alternatives such as GlutaMAX (L-alanyl-L-glutamine) for extended culture applications.
&nbsp;
<h2>What you will find:</h2>
&nbsp;
<ul>
<li>L-Glutamine stable formulations for sourcing of essential nitrogen</li>
<li>Non-Essential Amino Acids (NEAA) for growth media enhancement</li>
<li>Synthetic Polymer Coatings Poly-L-Ornithine and Poly-L-Lysine solutions for better cell attachment.</li>
<li>Extracellular Matrix Proteins used for cell adhesion and include high-purity Fibronectin and Vitronectin coating reagents</li>
<li>Adhesion Promotion Reagents for improving the surface properties of culture vessels.</li>
</ul>
&nbsp;
<h2>How to choose L-glutamine and amino acid supplements</h2>
&nbsp;
L-glutamine concentration and format
Most mammalian cell culture systems use L-glutamine concentrations between 2&ndash;6 mM, although requirements vary depending on the medium and cell type. It is commonly supplied as:
<ul>
<li>USP-grade powder for custom media preparation</li>
<li>200 mM liquid stock solutions for direct supplementation</li>
</ul>
Liquid stocks should be aliquoted and stored frozen to minimize repeated freeze-thaw cycles and preserve stability.
Understanding glutamine degradation
Free L-glutamine gradually breaks down in aqueous solution through a non-enzymatic process that produces ammonia and pyroglutamate. This degradation accelerates with increasing temperature, storage time, and exposure to common cell culture buffer systems.
Ammonia accumulation can negatively affect cell growth, viability, productivity, and experimental reproducibility, particularly during long culture periods without media replacement.
GlutaMAX and stable glutamine alternatives
GlutaMAX (L-alanyl-L-glutamine) is a stabilized dipeptide form of glutamine designed to resist degradation in culture media. Once taken up by cells, it is converted into usable glutamine while maintaining greater stability during incubation.
<ul>
<li>Reduced ammonia accumulation</li>
<li>Improved media stability</li>
<li>Better performance in long-term cultures</li>
<li>Less frequent media replacement requirements</li>
</ul>
For most applications, GlutaMAX can directly replace standard L-glutamine without protocol modifications.
MEM non-essential amino acids (NEAA)
NEAA supplements provide amino acids that cells can synthesize themselves but may benefit from receiving directly from the culture medium. Supplementation reduces the metabolic burden of amino acid production and often improves growth rates and cell viability.
NEAA solutions are typically supplied as 100X concentrates and diluted to 1X working concentration.
HT supplement
HT supplement contains hypoxanthine and thymidine and is commonly used following HAT selection during hybridoma production. It supports nucleotide synthesis through salvage pathways while cells recover from selection pressure.
MEM vitamin solution
Vitamin supplements provide additional growth-supporting micronutrients that may become limiting during prolonged culture or in demanding applications. These supplements are often used alongside amino acid supplementation to enhance culture performance.
&nbsp;
<h2>Specifications context</h2>
&nbsp;
The standard L-glutamine concentration used in many mammalian culture systems is approximately 2 mM, though actual requirements vary by medium formulation and cell type. Free L-glutamine remains stable as a dry powder and frozen stock solution but degrades progressively once added to liquid media.
Stabilized glutamine products such as GlutaMAX have become increasingly common because they maintain glutamine availability while minimizing ammonia accumulation during incubation. These formulations are particularly valuable in long-duration experiments and high-density culture systems.
<a href="https://mbpinc.net/contact-us">Contact the expert team</a> at MBP today to get research-grade supplements for your lab.

L-Glutamine & Amino Acids

L-Glutamine Solution (200 mM)

G275 by Applied Biological Materials (ABM) is a 200 mM L-Glutamine solution (29.2 g/L) in normal saline, 0.2 µm sterile-filtered. Supplied in 100 mL. Passes sterility testing. Store at −20°C; use within 2 weeks of thawing. For use as a cell culture media supplement.

Usage

Sterility

Storage Condition

<h2 data-section-id="185nm5c" data-start="0" data-end="38">What is classic cell culture media?</h2>
&nbsp;
Classic cell culture media refers to the foundational basal medium formulations that have underpinned mammalian cell culture for decades, including DMEM (Dulbecco's Modified Eagle Medium), MEM (Minimum Essential Medium), RPMI 1640, Ham's F-10, Ham's F-12, IMDM (Iscove's Modified Dulbecco's Medium), and DMEM/F-12. These media provide the amino acids, vitamins, glucose, salts, buffering agents, and other nutrients required to support cell survival and growth in vitro. They form the basis of many modern specialized, serum-free, and advanced media formulations used in academic research, biotechnology, pharmaceutical development, and biomanufacturing.
&nbsp;
<h2 dir="ltr">What you will find:</h2>
&nbsp;
<ul>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" aria-level="1">
&nbsp;Availability of both low and high carbohydrate levels of balanced basal formulations for the distinct metabolic requirements of diverse mammalian cell lines.
</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" aria-level="1">
Maintenance of precise physiological pH during long-term incubation cycles using optimized buffering configurations with integrated chemical stabilizers&nbsp;
</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" aria-level="1">
Nutrient-dense cell media supplemented with vitamins and essential amino acids to support&nbsp; hybridomas and rapidly proliferating continuous cultures
</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" aria-level="1">
Optimized serum reduction along with enhanced cell attachment using&nbsp;Multi-purpose 1:1 formulation hybrids&nbsp;
</li>
<li dir="ltr" style="font-family: arial, helvetica, sans-serif;" aria-level="1">
Salt solutions supplemented with essential minerals to facilitate enzyme preparation, cell washing, and reliable short-term osmotic maintenance.
</li>
</ul>
&nbsp;
<h2 data-section-id="1blio5n" data-start="931" data-end="964">How to choose a classic medium</h2>
&nbsp;
DMEM vs MEM
DMEM contains up to four times the amino acids and vitamins found in MEM, along with higher glucose concentrations, making it suitable for cell types with elevated nutrient demands. It is widely used for primary cells, stem cells, and many transformed mammalian cell lines. MEM, based on the original Eagle's formulation, remains a common choice for standard monolayer culture and is available with different salt formulations to support various applications.
RPMI 1640 for suspension and hematopoietic culture
RPMI 1640 was originally developed for culturing human leukemic cells and remains the preferred medium for lymphocytes, hybridomas, and other hematopoietic cell types. Its formulation includes glutathione and elevated levels of biotin, vitamin B12, and PABA. Although designed for suspension cultures, RPMI 1640 also supports a wide range of adherent cell lines when appropriately supplemented.
Ham's F-10/F-12 for CHO and specialized applications
Ham's nutrient mixtures were developed for CHO cells and other specialized cell types. Their formulations include additional nutrients such as zinc, putrescine, hypoxanthine, and thymidine, making them useful for hepatocyte culture, toxicity testing, viral fusion studies, and other specialized applications. These media can be used with or without serum depending on the experimental requirements.
DMEM/F-12 for complex, rich-medium applications
DMEM/F-12 combines the nutrient-rich composition of DMEM with the specialized components found in Ham's F-12. The resulting formulation supports demanding cell types including MDCK cells, glial cells, fibroblasts, and endothelial cells. It is also widely used as the foundation for many serum-free and advanced culture systems.
IMDM for high-density rapid proliferation
IMDM is an enriched derivative of DMEM that contains additional amino acids, vitamins, HEPES buffer, sodium pyruvate, and potassium nitrate. Its enhanced nutrient profile makes it particularly suitable for rapidly proliferating and high-density cultures such as Jurkat cells, COS-7 cells, and macrophages.
Liquid vs powder format
Liquid classic media are supplied ready for use after the addition of serum and required supplements. Powdered media offer lower shipping costs and greater flexibility for high-volume laboratories but require reconstitution, sodium bicarbonate addition, and sterile filtration before use.
&nbsp;
<h2 data-section-id="1fljvas" data-start="4382" data-end="4407">Specifications context</h2>
&nbsp;
Classic media are manufactured according to their original published formulations, with only stability-related modifications where necessary. Most liquid formulations contain sodium bicarbonate as the primary buffering system and require incubation in a 5&ndash;10% CO₂ atmosphere to maintain physiological pH. Powdered formulations require reconstitution, sodium bicarbonate supplementation, and sterile filtration before use.
As of 2026, DMEM, DMEM/F-12, and RPMI 1640 remain the most widely used classic cell culture media across academic, biotechnology, pharmaceutical, and biomanufacturing laboratories. IMDM continues to gain popularity for high-density culture and bioprocessing applications.
For recombinant growth factors and cytokines used alongside classic media, see Recombinant Proteins. For supplementation reagents such as GlutaMAX, NEAA, sodium pyruvate, and HEPES, see Cell Culture Supplements &amp; Additives. Browse Mammalian Cell Culture for the broader category.
<a href="https://mbpinc.net/contact-us">Contact the expert team</a> at MBP today and get optimized classic media for your lab.

Classic Media

M199 Medium

M199 Medium (TM502) by Applied Biological Materials is a 500 ml sterile basal medium containing L-glutamine, sodium bicarbonate (2.2 g/L), and phenol red (no HEPES) for broad-species primary cell culture, virology, and vaccine production. Supplement as required; requires 5–10% CO₂. Store at 2–8°C, protected from light.