BEWO Cell Line

BeWo is a human choriocarcinoma-derived cell line widely used in biomedical research. It serves as a valuable in vitro model for exploring the intricate process of placental development and syncytialization of human trophoblasts. Researchers also employ these cells to study trophoblast cell behaviour, hormone production, and substance transport across the placenta. Besides, BeWo cells also offer applications in toxicology, infection, and disease transmission studies.

This article encompasses fundamental knowledge about the BeWo cells that may help you deal with them in your research. It will include:

General characteristics and origin of BeWo cells
BeWo cell line: Culturing information
Advantages & disadvantages of BeWo cell line
Research applications of BeWo cells
Research Publications Featuring BeWo Cells
Resources for BeWo Cell line: Protocols, Videos, and More

1. General characteristics and origin of BeWo cells

Prior knowledge about a cell line's origin and general characteristics is crucial for its use in your research. This part of the article explores the BeWo cell line's origin, characteristic features, and much more. Here, you will learn: What is the BeWo choriocarcinoma cell line? What is BeWo syncytialization? From where do BeWo cells come from? What is the BeWo cell morphology? What is the ploidy of BeWo?

The placental cell line, BeWo, originated from the malignant gestational choriocarcinoma of a male fetal placenta in 1968 [1]. Thus, it serves as an invaluable model for placental research.
BeWo cells exhibit an epithelial-like morphology.
BeWo cells are relatively small, ranging between 20 to 30 µM [2].
BeWo cells exhibit a stable karyotype. Their stem line number is hypo-tetraploid. The modal chromosome number of BeWo cells is 86. It may vary from 71 to 178.
BeWo cells produce several hormones, i.e., human chorionic somatomammotropin, human chorionic gonadotropin (hCG), and steroid hormones.
BeWo cells express molecular markers, including CK 7, E-cadherin, and VE-cadherin.

B30 subline

B30 is the subline of BeWo cells. It is used to investigate the uptake and transport of nutrients. This is attributed to their vigorous growth on porous membranes.

BeWo and JEG-3

BeWo JEG-3 cell lines are derived from choriocarcinomas and used in placental research. BeWo cells produce hormones at lower levels than JEG-3, such as human chorionic gonadotropin (hCG) and estradiol.

Time-lapse visualization of human egg fertility, illustrating the stages of blastocyst formation, as observed under a microscope.

2. BeWo cell line: Culturing information

Maintaining a cell culture can be tricky unless you know key details. Such as: What is the doubling time of BeWo? What is BeWo cell media? How do you culture the BeWo cell line?, and How do you freeze BeWo cells?

Key Points for Culturing BeWo Cells

Doubling Time:	The population doubling time for the BeWo cells is approximately 30 hours.
Adherent or in Suspension:	BeWo is an adherent cell line.
Cell seeding density:	The seeding density recommended for BeWo cells is 1 x 10⁴ cells/cm². For seeding, old media is removed from cells, and cells are rinsed with 1 x PBS. Afterwards, Accutase (passaging solution) is added, and cells are incubated for 8 to 10 minutes at ambient temperature. Then, detached cells are added with culture media and centrifuged. The harvested cells are then resuspended and poured into a new flask.
Growth Medium:	Ham's F12K media containing 10% FBS, 2.0 mM L-Glutamine, 2.0 mM Sodium pyruvate, and 2.5 g/L NaHCO3 is used to grow BeWo cells. The BeWo cell media should be replaced 2 to 3 times per week.
Growth Conditions:	BeWo cells are maintained in a humidified incubator set at 37 degrees Celcius temperature and with a 5% CO2 supply.
Storage:	Frozen BeWo cells are stored at below -150°C in an electric ultra-low temperature freezer or in the vapour phase of liquid nitrogen.
Freezing Process and Medium:	BeWo cells are recommended to freeze in CM-1 or CM-ACF freezing medium. Cells are frozen using a slow freezing method. This method permits only a 1-degree decline in temperature per minute, thus preventing cells from any shock and protecting the viability of cells.
Thawing Process:	Frozen cells are thawed by immersing them in a pre-set 37°C water bath for 40 to 60 seconds until a small ice clump is left. Then, fresh growth media is added, and cells are centrifuged to remove the freezing media components. The collected cell pellet is resuspended and dispensed into a culture flask for growth.
Biosafety Level:	Biosafety Level 1 laboratory is necessary to handle and maintain BeWo cell cultures.

Capturing the epithelial, polygonal morphology, and islet-like growth of the human choriocarcinoma cell line, BEWO, through 10x and 20x magnification under the microscope.

3. Advantages & disadvantages of BeWo cell line

BeWo cells exhibit unique characteristics and come with specific pros and cons. In this section, we will explore some of the noteworthy ones.

Advantages

The main advantages of the BeWo choriocarcinoma cell line are:

In Vitro Placental Cell Model
BeWo cells provide an in vitro model for studying trophoblast cell behavior and placental syncytialization, replicating processes challenging to observe in vivo.
Hormone Production
BeWo cells secrete hormones, such as hCG and placental lactogen, making them valuable for hormonal regulation studies and research in reproductive biology.

Disadvantages

The disadvantages of BeWo trophoblast cells are:

Cancer cell line
BeWo cells were derived from choriocarcinoma, which is a placental tumor. This origin may not fully represent normal placental cell function.

4. Research applications of BeWo cells

The BeWo cell line is extensively used in placental research. A few specified applications of this cell line are mentioned here:

Placenta and pregnancy-related research: BeWo cells are pivotal in placental and pregnancy-related research. Such as researchers utilize these cells to study the transport of substances, including drugs, nutrients, and toxins, across the placental barrier. Besides, they also investigate the production and regulation of hormones, including human chorionic gonadotropin (hCG), placental lactogen, and steroid hormones. This is important for understanding hormonal changes during pregnancy. Research conducted in 2018 investigated the effect of brominated flame retardants (BFRs) on the activity of sulfotransferase in the human trophoblastic cell line (BeWo). This enzyme is involved in regulating the intracellular levels of the thyroid hormone [3].
Infection and Disease research: BeWo cells are also employed to examine the interaction between pathogens and placental cells. This research is valuable for understanding diseases that can be transmitted from mother to fetus and for developing strategies to protect fetal health. Moreover, this cell line helps researchers investigate complications that arise during pregnancy, such as preeclampsia. For instance, a study conducted by Michalina Bralewska in 2023 utilized the BeWo trophoblast cell model and found that both Chromogranin A (CgA) and the peptide derived from it, catestatin (CST), may have roles in the intricate process of preeclampsia (PE) pathogenesis [4]. Moving forward, another study explored the anti-cancer potential of trichosanthin using the BeWo choriocarcinoma cell line. The research also made an extra effort to discover biomarkers by performing comparative proteomic analysis before and after the experiment [5].

5. Research Publications Featuring BeWo Cells

Here are some significant research publications featuring the BeWo trophoblast cells.

AMPK Signaling Regulates Mitophagy and Mitochondrial ATP Production in Human Trophoblast Cell Line BeWo

This study in the Frontiers in Bioscience-Landmark (2022) proposed that AMPK signalling activates mitophagy in BeWo trophoblast cells via FUNDC1 and PRKN-facilitated mitophagy pathways. As a result, mitophagy helps maintain ATP production and mitochondrial membrane potential.

Ethylparaben induces apoptotic cell death in human placenta BeWo cells via the Caspase-3 pathway

This article was published in the Animal Cells and Systems (2020). The study proposed that ethylparaben mediates BeWo cells' apoptosis via regulating the caspase 3 pathway.

Characterization of Xenobiotic and Steroid Disposition Potential of Human Placental Tissue and Cell Lines (BeWo, JEG-3, JAR, and HTR-8/SVneo) by Quantitative Proteomics

This study in the drug Metabolism and Disposition (2023) utilized placental cell lines, including BeWo, JAR, JEG-3, and HTR-8/SVneo, to investigate the xenobiotics and steroid deposition in cells.

Apelin and apelin receptor in human placenta: Expression, signalling pathway and regulation of trophoblast JEG‑3 and BeWo cells proliferation and cell cycle

This research article in the International Journal of Molecular Medicine (2020) proposed that Apelin, the ligand of the APJ receptor, promotes BeWo cell proliferation through APJ and ERK1/2, stat3 and AMPK signaling. Thus, it could be an essential adipokine regulating the development of the placenta.

Antiseizure medications and fetal nutrients: effects on choline transporters in a human placental cell line

This study in the Epilepsia (2021) explored the effects of fetal nutrients and anti-seizures on choline transporters in the human-derived trophoblastic cell line, BeWo.

6. Resources for BeWo Cell line: Protocols, Videos, and More

Here are some online resources available on the BeWo cells:

In vitro transfection: This video will help you learn the in vitro plasmid DNA transfection protocol for adherent cells.

The following link contains cell culture information for BeWo cells:

BeWo cells: This website contains essential cell culture information about the BeWo cell line. This includes information regarding cell media and culturing conditions. It also has protocols for subculturing and handling cryopreserved and proliferative cultures.

References

Weber, M., et al., Cytogenomics of six human trophoblastic cell lines. Placenta, 2021. 103: p. 72-75.
Kolokol'tsova, T., et al., Characteristics of trophoblasts in long-term culture. Bulletin of Experimental Biology and Medicine, 2017. 164: p. 259-265.
Leonetti, C.P., C.M. Butt, and H.M. Stapleton, Disruption of thyroid hormone sulfotransferase activity by brominated flame retardant chemicals in the human choriocarcinoma placenta cell line, BeWo. Chemosphere, 2018. 197: p. 81-88.
Bralewska, M., T. Pietrucha, and A. Sakowicz, Reduction in CgA-Derived CST Protein Level in HTR-8/SVneo and BeWo Trophoblastic Cell Lines Caused by the Preeclamptic Environment. International Journal of Molecular Sciences, 2023. 24(8): p. 7124.
Hu, Y., et al., Comparative proteomic analysis of drug trichosanthin addition to BeWo cell line. Molecules, 2022. 27(5): p. 1603.