CLS will be called Cytion
Fastest deliveries on the market
> 800 well characterized cell lines
Worldwide service – one hand, one partner
Visit for your cell line needs

NCI-H1299 Cells: Research and Clinical Implications of NCI-H1299 Lung Cancer Cells

NCI-H1299 is an immortalized human non-small cell lung cancer cell line widely used in immune oncology, cancer, and drug discovery research. Moreover, researchers used these cells to investigate drug sensitivity, underlying signalling pathways, and molecular mechanisms related to lung cancer. Besides, these cells are employed to study viral infections such as SARS-CoV-2.

This article will discuss the fundamentals of NCI-H1299 cell lines that can help you learn all the necessary information and make your work easy. Mainly, you will know about the following:

  1. General characteristics and origin of NCI-H1299 cells
  2. NCI-H1299 cell line: Culturing information
  3. Advantages & disadvantages of NCI-H1299 cell line
  4. Research applications of NCI-H1299 cells
  5. Research Publications Featuring NCI-H1299 Cells
  6. Resources for NCI-H1299 Cell line: Protocols, Videos, and More

1.      General characteristics and origin of NCI-H1299 cells

The first thing to know about a cell line is its origin and general characteristics as they help you devise its use in your research. This section of the article will assist you in learning some important information about NCI-H1299's origin and characteristics. Such as, what are NCI-H1299 lung cancer cells? What type of cell is NCI-H1299? What is the size of NCI-H1299 cells? What is the difference between A549 and NCI-H1299?

  • The NCI-H1299 cell line originated from a lymph node metastasis of the lung of a 43-year-old Caucasian male patient with cancer. It was deposited by JD Minna and AF Gazdar.
  • These cells possess homozygous partial deletion of the p53 gene; thus, they do not express the p53 protein. Lack of NCI-H1299 p53 protein expression contributes to the proliferative tendency of H1299 lung cancer cells.
  • Besides p53, these immortalized cells are reported to have NCI-H1299 KRAS mutation, responsible for their growth, proliferation, migration, and invasion properties.
  • NCI-H1299 karyotype is near diploid.
  • NCI-H1299 cells have epithelial cell-like morphology.
  • These cells are flattened and have a thickness of less than 5 µm.

NCI-H1299 and A549

NCI-H1299 and A549 are non-small lung cancer cell lines. NCI-H1299 cells are more aggressive and sensitive compared to A549. Both have relatively similar mutations, such as KRAS. However, unlike NCI-H1299 cells, A549 cells express the P53 gene.

Animation of the SARS-Cov-2 virus in the human body attempting to enter a cell.

2.     Culturing information

Maintaining a cell line culture is not easy until you know all the key information about it, including: What is NCI-H1299 doubling time? What is the seeding density of NCI-H1299? What is NCI-H1299 cell media? How do you culture NCI-H1299 cell lines? This section will help you learn the answers to all these questions about NCI-H1299 cell culturing.

Key Points for Culturing NCI-H1299 Cells

Doubling Time:

NCI-H1299 doubling time ranges between 22-3o hours.

Adherent or in Suspension:

NCI-H1299 is an adherent cell line.

Sub-cultivation Ratio:

NCI-H1299 cells are sub-cultivated at a recommended ratio of 1:3 to 1:6. For seeding, adherent cells are washed with 1x PBS and incubated with Accutase passaging solution for 8 to 10 minutes at ambient temperature. Detached cells are added with fresh culture media and centrifuged. The cell pellet is then resuspended, and cells are poured into new flasks containing growth media.

Growth Medium:

RPM1 1640 is an ideal NCI-H1299 cell media. It is supplemented with 10% fetal bovine serum, 4500 mg/L glucose, 2.0 mM L-glutamine, 1 mM sodium pyruvate, 1500 mg/L NaHCO3, and 10 mM HEPES. Media should be renewed every 2 to 3 times a week.

Growth Conditions:

NCI-H1299 lung cancer cell cultures are maintained in a humidified incubator at 37°C with a 5% CO2 supply.


The NCI-H1299 cell line can be stored in the vapour phase of liquid nitrogen or ultra-low temperature freezers at below -150°C temperature for the long term.

Freezing Process and Medium:

CM-1 or CM-ACF are NCI-H1299 cells freezing media. Cells are frozen using a slow freezing process that permits only a 1-degree decrease in temperature per minute to protect cell viability.

Thawing Process:

Frozen NCI-H1299 cells are rapidly agitated in a preheated water bath at 37 degrees Celsius for 40 to 60 seconds until a small piece of ice is left. Thawed cells are added with fresh media and can be directly cultured in new flasks or centrifuged. In the former case, media should be replaced after 24 hours of incubation. Centrifugation helps to remove freezing media elements. Afterwards, harvested cells are resuspended in fresh medium and dispensed into the new flask for culturing.

Biosafety Level:

A biosafety level 1 laboratory is required to handle NCI-H1299 cell culture.


Adherent monolayer of NCI-H1299 at 20x and 40x magnification.

3.      Advantages & disadvantages of NCI-H1299 cells

Like other human cancer cell lines, NCI-H1299 also possesses some distinguishable characteristics associated with specific pros and cons. Herein, we have summarized a few significant ones.


The main advantages of the NCI-H1299 non-small cell lung cancer cell line are:

In Vitro lung cancer model

NCI-H1299 cell lines have been developed from lymph node metastasis of lung carcinoma and thus own relatively similar properties. Therefore, these cells can serve as an in vitro model to investigate lung cancer biology, cell and molecular mechanisms and screen and test potential therapeutics.

Easy to culture

NCI-H1299 cells are easy to culture and maintain in a research laboratory. There are no fussy cell culture requirements and procedures.

NCI-H1299 P53 and NCI-H1299 KRAS mutation

NCI-H1299 cells lack p53 gene expression, which allows cells to proliferate vigorously. Moreover, they possess KRAS mutation that contributes to cell growth, proliferation, invasion, and migration. Considering these, researchers use H1299 cells to investigate molecular mechanisms relevant to KRAS and P53 mutations.



The disadvantages of NCI-H1299 cells are:

Tissue specificity

NCI-H1299 originated from lung tissue. Therefore, its applicability is mostly limited to lung carcinoma research. It may not fully represent other cancer types' heterogeneity and general characteristics.


4.      Research applications of NCI-H1299 cells

The NCI-H1299 cell line is extensively used in lung cancer research. A few promising applications of this cell line are mentioned here.

  • Cancer biology: NCI-H1299 cells serve as a great research tool to investigate cancer development, progression, and related cell and molecular mechanisms. These cells equip certain mutations, allowing researchers to explore related cancer cell behaviours, signalling pathways, and gene expression profiles. Several studies have used NCI-H1299 non-small cell lung carcinoma cell line to study lung cancer biology. For instance, a study conducted in 2018 investigated the signalling pathway involved in lung cancer cell apoptosis using NCI-H1299 cells. The researchers found that PI3K/AKT pathway contributes to cell proliferation, and its inhibition can cause cancer cell death [1]. Likewise, another research investigated the mechanisms involved in NCI-H1299 epithelial-to-mesenchymal transition (EMT). This study proposed that an extracellular protein SPARC (secreted protein acidic and rich in cysteine) contributes to NCI-H1299 EMT and migration, thus, promotes tumorigenesis in cells—the SPARC protein act as a mediator of TGF-β1 signalling [2]. 
  • Drug discovery and screening: NCI-H1299, a human lung cancer cell line, is extensively employed to assess the toxicity and efficacy of potential drugs. In addition, it is used by researchers to investigate the mechanism of action of anti-tumour drugs. Studies have also used these cells to identify possible drug targets and resistance mechanisms. For example, research conducted by Xiao-Yun Shen and colleagues investigated the anti-cancer potential of the natural compound bruceine D (BD). The findings revealed that BD significantly impedes the proliferation, invasion, and migration of NCI-H1299 cells. Therefore, it may be considered a potential adjuvant therapy for treating non-small cell lung carcinoma [3]. Similarly, a study evaluated the cytotoxic effects of xanthohumol alone and in combination with cisplatin chemotherapeutic drug on NCI-H1299 lung cancer cells [4].

5.      Research Publications Featuring NCI-H1299 Cells

Some significant and most cited research publications about the NCI-H1299 lung carcinoma cell line are here.

Induction of apoptosis in indole-3-carbinol-treated lung cancer H1299 cells via ROS level elevation

This article is published in the Human and Experimental Toxicology journal (2021). The study proposed that indole-3-carbinol induces apoptosis in H1299 via elevating reactive oxygen species (ROS) levels.

Synergistic anticancer activity of combined use of caffeic acid with paclitaxel enhances apoptosis of non-small-cell lung cancer H1299 cells in vivo and in vitro

This research was published in 2018 in Cellular Physiology and Biochemistry journal. The research findings revealed that caffeic acid and paclitaxel combination exert a synergistic anti-cancer effect in non-small cell lung cancer cell line, NCI-H1299.

Antitumor effects of Tubeimoside-1 in NCI-H1299 cells are mediated by microRNA-126-5p-induced inactivation of VEGF-A/VEGFR-2/ERK signaling pathway

This research article was published in Molecular Medicine Reports (2018). The study evaluated the anti-tumor effect of tubeimoside-1, a triterpenoid saponin, and underlying mechanisms in the NCI-H1299 cell line.

Pristimerin induces apoptosis and inhibits proliferation, migration in H1299 lung cancer cells

This article published in the Journal of Cancer (2020) proposed that Pristimerin, a natural compound suppresses the proliferation and migration of NCI-H1299 cells.

Mechanism of VIPR1 gene regulating human lung adenocarcinoma H1299 cells

This study in Medical Oncology (2019) explored the potential effects of VIPR1 gene overexpression on the NCI-H1299 adenocarcinoma cell line.

6.      Resources for NCI-H1299 Cell line: Protocols, Videos, and More

Here are some online resources featuring the NCI-H1299 cells.

The following link contains cell culture information for NCI-H1299 cells. 

  • NCI-H1299 cell line: This link can help you know about the NCI-H1299 cell passaging and transfection protocol.


  1. Gu, J., et al., Study of EGCG induced apoptosis in lung cancer cells by inhibiting PI3K/Akt signaling pathway. Eur. Rev. Med. Pharmacol. Sci, 2018. 22(14): p. 4557-4563.
  2. Sun, W., et al., SPARC acts as a mediator of TGF‐β1 in promoting epithelial‐to‐mesenchymal transition in A549 and H1299 lung cancer cells. Biofactors, 2018. 44(5): p. 453-464.
  3. Shen, X.-Y., et al., A study on the mechanism of bruceine D in the treatment of non-small cell lung cancer H1299 cells. World Journal of Traditional Chinese Medicine, 2020. 6(4): p. 500.
  4. Long, B., et al., Cytotoxic Effects of Xanthohumol and Its Combination with Cisplatin on Human Metastatic Lung Cancer H1299 Cells. Journal of Advances in Medicine and Medical Research, 2019. 30(9): p. 1-15.