Written and maintained by CASRAI Editorial Board
Last updated
The HT-29 cell line (also written HT29) is a human colorectal adenocarcinoma line that grows as adherent epithelial cells. It was isolated in 1964 from the primary tumor of a 44-year-old woman and is catalogued by ATCC as HTB-38. A cell line, in the broad sense used by Cellosaurus, is a population of cells from a tissue that can be propagated and shared between laboratories as a reusable research model.
Definition and scope: what is a cell line?
A cell line is a population of cultured cells that can be kept in the laboratory and distributed for repeated experiments. Cellosaurus, the knowledge resource on cell lines run by the SIB Swiss Institute of Bioinformatics, describes its scope as immortalized cell lines, naturally immortal lines such as stem cell lines, and finite-life lines that are widely distributed. It covers vertebrate (with emphasis on human, mouse and rat), insect, tick and plant lines, and it excludes primary cell lines other than those finite-life exceptions. That is the working definition behind the related searches “cell line definition”, “definition of cell line” and “what is a cell line”.
The plural, “cell lines”, matters in practice because every line has an identity record: a recommended name, synonyms, a unique accession number, species, tissue of origin and notes on misidentification or misspellings. For HT-29 the Cellosaurus accession is CVCL_0320, and the recorded synonyms are “HT 29” and “HT29”.
What is the HT-29 cell line?
ATCC describes HT-29 as a human colon, colorectal adenocarcinoma line, an adherent epithelial cell line with enterocyte-like features. J. Fogh isolated it in 1964 from a primary tumor by explant culture; the donor was a 44-year-old White female. Cellosaurus lists the originating institution as Memorial Sloan Kettering Cancer Center and records the same age and sex.
Characteristics reported by ATCC and Cellosaurus
- Morphology: epithelial; the cells form a tight monolayer with an apical brush border (ATCC).
- Karyotype: hypertriploid, with a modal chromosome number of 71 and a range of 68 to 72 (ATCC).
- Tumorigenicity: tumorigenic in nude mice, producing a grade I, well-differentiated adenocarcinoma (ATCC).
- Microsatellite status: microsatellite stable (MSS) (Cellosaurus).
- Recorded mutations: APC (heterozygous), BRAF p.Val600Glu (heterozygous), PIK3CA p.Pro449Thr (heterozygous), SMAD4 p.Gln311Ter (homozygous) and TP53 p.Arg273His (homozygous) (Cellosaurus).
- Listed applications: cancer research, toxicology, high-throughput screening, gastroenterology, 3D culture, use as a transfection host, and absorption, transport and secretion studies as a gastrointestinal barrier model (ATCC).
How HT-29 is cultured in practice
ATCC specifies the following conditions for HTB-38. They are product specifications from the supplier, not a bench protocol, and a laboratory should work from its own SOP and the lot documentation.
| Item | ATCC HTB-38 specification |
|---|---|
| Growth medium | ATCC-formulated McCoy’s 5A Modified Medium (30-2007) with fetal bovine serum to a final concentration of 10% |
| Atmosphere and temperature | 95% air, 5% CO2; 37 C |
| Subculture ratio | 1:3 to 1:8 |
| Medium renewal | 2 to 3 times per week |
| Freeze medium | Complete growth medium with 5% (v/v) DMSO |
| Storage | Vapor phase of liquid nitrogen; ATCC warns that storage at -70 C will result in loss of viability |
For anyone assembling consumables for this work, the table above is the sourced list of what the supplier specifies: a defined basal medium, a serum concentration, a freezing additive and ultra-cold storage. Pack sizes, brands and quantities are not stated by ATCC and are not covered here.
Related cell lines
HT-29 is one of several widely used cancer and epithelial models. CASRAI has separate explainers for the HCT116 cell line, the MCF7 cell line, the HepG2 cell line and the origin of HeLa cells. Intestinal barrier work is covered in the guide to the Caco-2 permeability assay. Cellosaurus also records derivative lines related to HT-29, including WiDr, KM20 and drug-resistant or knockout sublines such as HT29-MTX, which is why the exact line name on a vial label matters.
Identity checks for HT-29 on receipt
Because look-alike and derivative names exist, the useful question on receipt is whether the vial is what the label says. These items come from ATCC and Cellosaurus.
- Name: confirm the name and accession (HTB-38 at ATCC, CVCL_0320 at Cellosaurus) rather than a shortened “HT29” alone.
- Amelogenin and STR profile: ATCC lists Amelogenin X, CSF1PO 11,12, D13S317 11,12, D16S539 11,12, D5S818 11,12, D7S820 10, TH01 6,9, TPOX 8,9 and vWA 17,19 (selected loci). A profile that departs from these needs an explanation before experiments proceed.
- Contamination status: ATCC reports bacteria, fungi and mycoplasma as not detected for its lot; a laboratory that expands the line should keep its own testing records.
- Karyotype: a hypertriploid modal number near 71 is the ATCC reference.
Reported doubling times differ by source
ATCC does not state a doubling time. Cellosaurus records values from several sources (a selection is shown), and they do not agree, so no single number should be used for scheduling.
| Source recorded in Cellosaurus | Doubling time |
|---|---|
| NCI-DCTD | 19.5 hours |
| CLS (300215) | about 24 hours |
| PubMed 3281752 | 26.9 +/- 3.3 hours |
| PubMed 25944804 | 36.93 hours |
| DSMZ (ACC-299) | about 40 to 60 hours |
Where it matters for labs and supply decisions
Cellosaurus notes that HT-29 belongs to several reference panels, including the NCI-60 cancer cell line panel, the Cancer Dependency Map project (which includes the Cancer Cell Line Encyclopedia), the COSMIC cell lines project and the MD Anderson Cell Lines Project. A laboratory comparing its results with published screens therefore needs the same line identity. The ATCC conditions in the culture table are also the reference when a laboratory writes consumable lists, such as freezing vials, DMSO and serum, for a new project.
Common misunderstandings
“HT29”, “HT-29” and “HCT29” are the same thing
HT 29, HT29 and HT-29 are recorded synonyms. HCT29 is recorded in Cellosaurus as a misspelling, and it is easily confused with HCT116, a different line.
HT-29 always means colon
The oldest report calls the tissue colon, and Cellosaurus notes that one publication describes it as rectosigmoid adenocarcinoma. ATCC uses colon, colorectal adenocarcinoma.
Biosafety level
ATCC’s HTB-38 page carries inconsistent biosafety wording, so the level to apply should be confirmed with ATCC’s current documentation and the institutional biosafety committee. This guide does not state a biosafety level.
FAQ
What is the HT29 cell line used for?
ATCC lists cancer research, toxicology, high-throughput screening, gastroenterology, 3D culture, transfection and absorption, transport and secretion studies.
Where does the HT-29 cell line come from?
A primary colorectal adenocarcinoma from a 44-year-old woman, isolated in 1964 by J. Fogh using explant culture.
Is HT-29 adherent?
Yes. ATCC calls it an adherent epithelial cell line that forms a tight monolayer with an apical brush border.
How fast does HT-29 grow?
Sources disagree: Cellosaurus lists values from 19.5 hours to about 40 to 60 hours, depending on the source.
How should HT-29 cells be stored?
ATCC specifies the vapor phase of liquid nitrogen with a 5% DMSO freeze medium, and warns that storage at -70 C will result in loss of viability.
Sources
- ATCC, HT-29 (HTB-38) product page, https://www.atcc.org/products/htb-38, checked 2026-10-29.
- Cellosaurus, HT-29 entry (CVCL_0320), https://www.cellosaurus.org/CVCL_0320, checked 2026-10-29.
- Cellosaurus, Description of the resource, https://www.cellosaurus.org/description.html, checked 2026-10-29.
