HT-29 Cells
















Overview of the HT-29 cell line
Description | The HT-29 cell line, derived from a Grade II human colorectal adenocarcinoma, represents a cornerstone research model in the study of human colon cancers. Derived from a primary tumor in a 44-year-old female in 1964, HT22 cells have been instrumental in advancing our understanding of the adhesion or invasion mechanisms of cancer cells. As a human adenocarcinoma cell line, HT-29 cells exhibit characteristics that closely mimic those of mature intestinal cells, such as enterocytes, underscoring their utility in exploring the dynamics of food digestion and nutrient bioavailability. HT-29 cells are sensitive to conventional colorectal cancer chemotherapies, including 5-fluorouracil and oxaliplatin. This sensitivity, coupled with their ability to express differentiation pathways under specific conditions, such as glucose deprivation or treatment with inducers like butyrate, makes them an invaluable model for investigating the molecular mechanisms underlying cell differentiation and cancer progression. Moreover, HT-29 cells have been utilized as a xenograft tumor model, providing a platform for in vivo studies that mimic the tumor's behavior in the human body. This application allows for the exploration of tumor growth, metastasis, and the efficacy of therapeutic agents in in vivo situations. In summary, the HT-29 cell line is a pivotal tool in medical and biological research, facilitating a deeper understanding of human colon adenocarcinoma, the molecular basis of cancer cell differentiation, and the development of effective cancer treatments. |
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Organism | Human |
Tissue | Colon |
Disease | Adenocarcinoma |
Synonyms | HT 29, HT29 |
Specifications
Age | 44 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Epithelial-like |
Growth properties | Adherent |
Documentation
Citation | HT-29 (Cytion catalog number 300215) |
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Biosafety level | 1 |
NCBI_TaxID | 9606 |
CellosaurusAccession | CVCL_0320 |
Genomics of the colorectal adenocarcinoma cell line HT29
Receptors expressed | urokinase receptor(u-PAR), vitamin D (moderate expression), no detectable plasminogen activator activity. |
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Protein expression | CEA negative, p53 positive |
Antigen expression | Blood Type A, Rh+, HLA A1, A3, B12, B17, Cw5, CD4 -, cell surface expression of galactose ceramide (a possible alternative receptor for HIV) |
Isoenzymes | Me-2, 1, PGM3, 1-2, PGM1, 1-2, ES-D, 1, AK-1, 1, GLO-1, 1-2, G6PD, B, Phenotype Frequency Product: 0.0230 |
Oncogenes | myc+, ras+, myb+, fos+, sis+, p53+, abl -, ros -, src - |
Tumorigenic | Yes, in nude mice. Forms well differentiated adenocarcinoma consistent with colonic primary (grade I), tumors also form in steroid treated hamsters |
Virus susceptibility | human immunodeficiency virus (HIV, LAV) |
Products | Secretory component of IgA, carcinoembryonic antigen (CEA), transforming growth factor beta binding protein, mucin, The p53 antigen is overproduced |
Karyotype | The stemline chromosome number is hypertriploid with the 2S component occurring at 2.4%. Seventeen marker chromosomes are found in most metaphases, generally in single copy per chromosome. The marker designations are: M1p-(=t(3p-,?) with a deleted short arm), t(7q,?), t(10q,?), i(13q), 19q+a. M6, ?t(8q,9q-), ?xp, M9, 6q+, t(13,?)a, t(13,?)b, 19q+b, M14, M15, 15p+, and xq-. Chromosome 13 is nullisomic and chromosomes 8 and 14 are generally monosomic. No Y chromosome was detected by QM band analysis. |
Handling
Culture Medium | EMEM (MEM Eagle), w: 2 mM L-Glutamine, w: 2.2 g/L NaHCO3, w: EBSS (Cytion article number 820100a) |
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Supplements | Supplement the medium with 10% FBS and 1% NEAA |
Dissociation Reagent | Accutase |
Doubling time | 24 hours |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:3 to 1:8 is recommended |
Seeding density | 3 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Post-Thaw Recovery | Slow, the cells need roughly 48 hours to settle and adhere. |
Freeze medium | As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress. |
Thawing and Culturing Cells |
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Quality Verification
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
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STR profile |
Amelogenin: x,x
CSF1PO: 11,12
D13S317: 11
D16S539: 11,12
D5S818: 11,12
D7S820: 10
TH01: 6,9
TPOX: 8,9
vWA: 17,19
D3S1358: 15,17
D21S11: 29,3
D18S51: 13
Penta E: 14,16
Penta D: 11,13
D8S1179: 10
FGA: 20,22
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HLA alleles |
A*: '01:01:01, '24:03:01
B*: '35:01:01, '44:03:01
C*: '04:01:01
DRB1*: '04:02:01, '07:01:01
DQA1*: '02:01:01, '03:01:01
DQB1*: '02:02:01, '03:02:01
DPB1*: '04:01:01
E: '01:01, '01:03
|
Required products
What's in EMEM?
EMEM is a modified version of Eagle's minimum essential medium, containing Earle's Balanced Salt Solution, non-essential amino acids, L-glutamine, sodium pyruvate, and sodium bicarbonate. It's important to note that this level of sodium bicarbonate is intended for use in 5% CO2 in the air. To maintain its effectiveness, storing the medium at 2°C to 8°C in the dark when not in use is recommended.
What is EMEM used for?
Eagle's minimal essential medium (EMEM) is a cell culture medium that can maintain cells in tissue culture. The medium contains higher concentrations of amino acids, allowing for a more accurate approximation of the protein composition of cultured mammalian cells. EMEM may be used to cultivate various cells, including fibroblasts, human liver cancer cell line (HepG2) cells and human fetal brain progenitor-derived astrocyte cells (PDA). It is typically used in the presence of fetal bovine serum (FBS), calf, or horse sera.
How is EMEM different from other cell culture media?
While EMEM and Dulbecco's modified Eagle's medium (DMEM) share some similarities, they also differ. Both media lack protein and contain the amino acids, salts, glucose, and vitamins required to provide a cell with energy and maintain it in tissue culture. However, the DMEM formulation is modified to contain up to four times more vitamins and amino acids and two to four times more glucose than EMEM. It's worth noting that EMEM is also different from the original MEM formulation.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +8°C, protected from light.
Once opened, store at 4°C and use within 6–8 weeks.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Amino Acids
L-Arginine HCl
126.00
L-Cystine 2 HCl
31.30
L-Glutamine
292.00
L-Histidine HCl H2O
42.00
L-Isoleucine
52.00
L-Leucine
52.00
L-Lysine HCl
72.50
L-Methionine
15.00
L-Phenylalanine
32.00
L-Threonine
48.00
L-Tryptophan
10.00
L-Tyrosine 2 Na 2 H2O
51.90
L-Valine
46.00
Vitamins
Choline Chloride
1.00
Vitamins
D-Calcium Pantothenate
1.00
Folic Acid
1.00
myo-Inositol
2.00
Nicotinamide
1.00
Pyridoxal HCl
1.00
Riboflavin
0.10
Thiamine HCl
1.00
Inorganic Salts
CaCl2 2 H2O
265.00
Inorganic Salts
KCl
400.00
MgSO4
97.67
NaCl
6800.00
NaHCO3
2200.00
NaH2PO4
122.00
Other Components
D-Glucose
1000.00
Other Components
Phenol Red Sodium Salt
11.00
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a widely used buffer solution in biological and chemical research. It plays a crucial role in maintaining the pH balance and osmolarity during various experimental procedures, including tissue processing and cell culture. Our PBS solution is meticulously formulated with high-purity ingredients to ensure stability and reliability in every experiment. The osmolarity and ion concentrations of our PBS closely mimic those of the human body, making it isotonic and non-toxic to most cells.
Composition of Our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains:
8000 mg/L Sodium chloride (NaCl)
200 mg/L Potassium chloride (KCl)
1150 mg/L Sodium phosphate dibasic anhydrous (Na2HPO4)
200 mg/L Potassium phosphate monobasic anhydrous (KH2PO4)
This composition ensures an optimal pH and ionic balance, suitable for a wide range of biological applications.
Applications of Our PBS Solution
Our PBS solution is ideal for various applications in biological research. Its isotonic and non-toxic properties make it suitable for substance dilution and cell container rinsing. PBS solutions containing EDTA are effective for disengaging attached and clumped cells. However, divalent metals such as zinc should not be added to PBS, as this can cause precipitation. In such cases, Good's buffers are recommended. Additionally, our PBS solution is an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, including SARS-CoV-2.
Quality Control
Sterile-filtered
Storage and Shelf Life
Store at +2°C to +25°C, protected from light.
Once opened, store at 2°C to 25°C and use within 24 months.
Shipping Conditions
Ambient temperature
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Avoid freezing and frequent warming to +37°C, as it reduces product quality.
Do not heat the medium beyond 37°C or use uncontrolled heat sources such as microwave appliances.
If only part of the medium is to be used, remove the required amount and warm it to room temperature before use.
Composition
Category
Components
Concentration (mg/L)
Salts
Potassium chloride
200
Potassium phosphate monobasic anhydrous
200
Sodium chloride
8000
Sodium phosphate dibasic anhydrous
1150
Related products
Certificate of Analysis (CoA)
Lot Number | Certificate Type | Date | Catalog Number |
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300215-140225 | Certificate of Analysis | 23. May. 2025 | 300215 |
300215-921 | Certificate of Analysis | 23. May. 2025 | 300215 |
Material Transfer Agreement
If you intend to use Cytion cell lines solely for internal research at a single research site, please complete and sign our Material Transfer Agreement (MTA) and submit it along with your order.
For any commercial applications - including but not limited to fee-for-service work, quality control testing, product release, diagnostic use, or regulatory studies - please complete the Intended Use Form so we can prepare a suitable agreement tailored to your project.
Please note: The MTA applies only to certain cell lines. If this notice and the MTA document appear on a product page, the agreement is applicable. For cell lines not covered by the MTA, no reference to the agreement will be shown. The MTA is not valid for customers in the Americas, China, or Taiwan. Please contact our U.S. entity to receive the appropriate agreement.