HL-60 Cells: Acute Myeloid Leukemia Research Essentials

HL-60 is an immortal human leukemia cell line. It is widely used to study cancer biology and to explore the efficacy of anti-cancer drugs. Moreover, it can also be used to understand different cellular and molecular mechanisms involved in leukemic cell proliferation and differentiation.

Before working on any cell line, you should know a few key points. In this article, you will learn about the HL-60 cell line.

HL-60 cell line: Origin and general information
HL-60 cell culture information
HL-60 cells: Advantages and disadvantages
Applications of HL-60 cell line in research
HL-60 cells: Research publications
HL-60 cell line Resources: Protocols, Videos & More

1. HL-60 cell line: Origin and general information

Whenever we talk about a cell line, we first think about its origin and distinguishing features. In this section, you will learn: What is the origin of HL-60 cells? What is the morphology of the HL-60 cell line? What are the characteristics of HL-60 cells?

What is the HL-60 leukemia cell line?

HL-60 is a human leukemia cell line derived from a 36-year-old female patient (Caucasian) having acute promyelocytic leukemia. These were the first myeloid leukemia cells successfully isolated and cultured in suspension form by S.J. Collins and his colleagues [1].

Morphology of HL-60 cells

HL-60 possesses a lymphoblastic morphology. These cells grow in suspension cultures, so they are round-shaped.

Cell size

The HL-60 cells are heterogeneous in size; therefore, their diameter ranges from 9 to 25 μm [1].

Genome and ploidy

The modal chromosome number reported for HL-60 is 43. The stemline chromosome number of the HL-60 cell line is pseudodiploid , with the occurrence of a 2S component at a 6.2 % rate.

Monocellular leukemia tissue under the microscope.

2. HL-60 cell culture information

HL-60 cell culturing is simple and easy. This section will familiarize you with basics such as What is the doubling time of the HL-60 cell line? Is HL-60 adherent or suspension culture? What is the seeding density of the HL-60 cell line? What medium is used for HL-60 growth?

Doubling time:	The doubling time for HL-60 cells is approximately 36 to 48 hours.
Adherent or in suspension:	HL-60 cells grow as suspension cultures.
Seeding density:	These cells are seeded at 1 x 10⁶ cells/ml density when subcultured and at 2 x 10⁵ cells/ml when a new culture is started.
Growth medium:	RPMI 1640 growth medium added with 10 % FBS and 2.5 mM L-glutamine is used to culture HL-60 cells. The medium should be renewed 2 to 3 times per week.
Growth conditions:	HL-60 cells are grown in a humidified incubator at 37 °C temperature and 5 % CO₂ supply.
Storage:	HL-60 cells are stored at below -150 °C temperature or in the vapour phase of liquid nitrogen to keep a maximum number of cells viable.
Freezing medium and process:	CM-1 or CM-ACF freezing media are used for HL-60 cell freezing. A slow freezing method is preferred for the HL-60 cell freezing.
Thawing process:	Frozen vial of HL-60 cells is rapidly agitated in a water bath set at 37 °C temperature. Afterwards, cells are centrifuged to remove freezing media. New culture media is added, and cells are resuspended to dispense into a flask for growth.
Biosafety level:	Biosafety level 1 is recommended for handling HL-60 cultures.

HL-60 suspension cells at 10x and 20x magnification.

3. HL-60 cells: Advantages and disadvantages

Before working with any cell line, you might want to know about its advantages and drawbacks. This article section will cover a few pros and cons of using HL-60 cells.

Advantages

A few prominent advantages of the HL-60 cell line are:

Versatility	HL-60 cells have the ability to differentiate into multiple cell types, including monocytes, macrophages, and granulocytes, under appropriate growth conditions. Thus they can be used to study cellular differentiation and other cell processes.
Easy to culture	HL-60 cells are easy to grow and maintain in suspension cultures. Therefore, they are a convenient tool for studying multiple cell processes.
In vitro model of leukemia	These cells were obtained from a patient with acute myeloid leukemia, and thus have genetic and molecular characteristics similar to the disease.

Disadvantages

The disadvantages of the HL-60 cell line are:

Bacterial contamination	Like all human cell lines, HL-60 cells are also susceptible to contamination by bacteria, fungi, and other microbes. Contamination can be avoided by maintaining aseptic culture conditions.
Limited differentiation potential	HL-60 can differentiate into some myeloid lineages in response to stimuli. The differentiation potential of HL-60 cells is limited compared to the primary cells.

4. Applications of HL-60 cell line in research

HL-60 are low-maintenance cultures that have many research applications. Here, we will discuss a few significant ones.

Cancer Research: HL-60 cell line is extensively used in cancer research. These cells are used to study cancer cell signalling, identify drug targets, and screening of new chemotherapeutic agents. A study used HL-60 cells to investigate the anti-cancer potential of Stigmast-5-en-3-ol, a chemical extracted from Dendronephthya gigantea named soft corals. This study indicated that the chemical exerts antiproliferative and apoptotic activity against human leukemia cell line HL-60 [2].

Another research examined the role of the JAK/STAT signalling pathway in the proliferation of HL-60 leukemia cells. This study also utilized HL-60 cells to investigate the potential therapeutic effect of a compound, Thymoquinone [3].

Immunology: The HL-60 cell line has been utilized to investigate different immune cell responses, including inflammation. To study these processes, these cells are differentiated into macrophage, monocyte, and neutrophil-like cells. A recent study used HL-60 cells to develop a macrophage-like cell model and studied the anti-inflammatory potential of Brassica plant constituents [4]. Another study experimentally developed neutrophil-like cells from the HL-60 leukemia cell line to investigate the genes regulating the cell differentiation processes [5].
Drug Discovery: HL-60 cells have been used as a screening tool to identify potential drug candidates for cancer treatment. Such as, a study used HL-60 leukemia cells to investigate the potential anticancer activities of Brassica plant compounds [6].

5. HL-60 cells: Research publications

Several research studies are available on HL-60 cells. Here, we have listed some notable examples:

Apoptotic and antiproliferative effects of Stigmast-5-en-3-ol from Dendronephthya gigantea on human leukemia HL-60 and human breast cancer MCF-7 cells

This research paper proposed the anti-tumor effect of a compound, Stigmast-5-en-3-ol, on HL-60 leukemia cells and MCF7 breast cancer cells.

Thymoquinone suppresses cell proliferation and enhances apoptosis of HL60 leukemia cells through re-expression of JAK/STAT negative regulators

This study describes the role of JAK/STAT signalling in the proliferation of HL-60 cells. In addition, researchers investigate the therapeutic potential of the Thymoquinone compound using this leukemia cell line. The compound inhibits cell proliferation and induces cell death by reviving the expression of negative regulators of the JAK/STAT cascade.

Analysis of the anti-inflammatory potential of Brassica bioactive compounds in a human macrophage-like cell model derived from HL-60 cells

This study utilized HL-60 cells to derive a macrophage-like cell model from investigating the anti-inflammatory activity of Brassica plant compounds.

ERK is involved in the differentiation and function of dimethyl sulfoxide-induced HL-60 neutrophil-like cells, which mimic inflammatory neutrophils

This study published in the International Immunopharmacology journal developed neutrophil cell mimic from the HL-60 cell line by using dimethyl sulfoxide. The study observed that the ERK gene is involved in differentiating these cells.

Antitumour effects of selected plant polyphenols, gallic acid and ellagic acid, on sensitive and multidrug‐resistant leukaemia HL60 cells

This research paper investigates the anti-cancer potential of plant polyphenolic compounds, i.e., gallic acid and ellagic acid, on drug-sensitive and resistant HL-60 leukemia cells.

6. HL-60 cell line Resources: Protocols, Videos & More

Multiple online educational resources are available on the HL-60 cell line. This section will cover a few resources describing the culturing and transfection of this cell line.

Cell culture protocols

The resources describing HL-60 cell culturing methods are:

Propagation of HL-60: This document will provide information about subculturing of HL-60 cells.
HL-60 cell line: This website contains the basic information on HL-60 cells and protocols, including culturing, freezing, and thawing of cells.

Transfection protocols

Here are some resources describing transfection methods opted for HL-60 cells.

Transfection of HL-60 cells: This link has a detailed transfection procedure for HL-60 leukemia cells.
HL-60 cells transient transfection: This document contains useful information about the cell line and gives a detailed protocol for transient transfection of HL-60 cells.

Videos related to HL-60 cell line

There are very limited video resources on HL-60 cells.

Passaging HL-60 cells: This video explains the protocol for subculturing HL-60 cells.

We expect this article to provide you with basic information about culturing and maintaining the HL-60 cell line. In addition, it can enhance your knowledge about the advantages of using these leukemia cells in research. If you are planning on using this cell line, order from us.

References

Fleck, R., S. Romero-Steiner, and M. Nahm, Use of HL-60 cell line to measure opsonic capacity of pneumococcal antibodies. Clinical and Vaccine Immunology, 2005. 12(1): p. 19-27.
Fernando, I.S., et al., Apoptotic and antiproliferative effects of Stigmast-5-en-3-ol from Dendronephthya gigantea on human leukemia HL-60 and human breast cancer MCF-7 cells. Toxicology in Vitro, 2018. 52: p. 297-305.
Almajali, B., et al., Thymoquinone suppresses cell proliferation and enhances apoptosis of HL60 leukemia cells through re-expression of JAK/STAT negative regulators. Asian Pacific Journal of Cancer Prevention: APJCP, 2021. 22(3): p. 879.
Ruiz-Alcaraz, A.J., et al., Analysis of the anti-inflammatory potential of Brassica bioactive compounds in a human macrophage-like cell model derived from HL-60 cells. Biomedicine & Pharmacotherapy, 2022. 149: p. 112804.
Wang, D., et al., ERK is involved in the differentiation and function of dimethyl sulfoxide-induced HL-60 neutrophil-like cells, which mimic inflammatory neutrophils. International Immunopharmacology, 2020. 84: p. 106510.
Núñez-Sánchez, M.Á., et al., Anti-Leukemic Activity of Brassica-Derived Bioactive Compounds in HL-60 Myeloid Leukemia Cells. International Journal of Molecular Sciences, 2022. 23(21): p. 13400.