Two new articles published in Nature are the focus of a new Scientific American article: “Mini-Me Brains Mimic Disease, Raise Hope for Eventual Therapies”.  One article, “Assembly of Functionally Integrated Human Forebrain Spheroids” focuses on the creation of a new disease model where brain organoids were generated from patients with Timothy syndrome.  The other article, “Cell Diversity and Network Dynamics in Photosensitive Human Brain Organoids” describes generation of brain organoids that “develop spontaneous networks and photosensitive neurons that can be modulated by sensory stimulation with light.”

Several researchers who were not a part of the study were interviewed for comment, including Dr. Juergen Knoblich, (whose lab generated the first cerebral organoids in 2013) and Dr. Timothy O’Brien, (who used Cell-Mate3D to generate complex cerebral organoids using only the matrix and iPSC maintenance media.)

Figure 1. Cerebral organoid developed in Dr. Timothy O’Brien’s laboratory containing neural tube-like structures. Nestin (green) Sox1 (red). Courtesy of Dr. Timothy O’Brien at the University of Minnesota.

Dr. Knoblish commented on the articles:

“This shows that the approach has much greater potential than we ever imagined.” “They’ve shown that if you keep [the mini-brain] growing for a long enough time, it will generate the whole repertoire of cells we see in the human brain.”

The ethics of mini-brains was also discussed.  Dr. O’Brien stated:

“The largest mini-brains-in-a-dish are only 4 millimeters across — roughly the size of a sea slug or jellyfish brain” — and, “a tiny, tiny fraction of the human brain”. “You do see some neural circuits forming, but none that are anywhere near the size needed for sentience, and they are not nearly complicated enough to feel pain.”  “Developing better mimics of the human brain will take a lot more time, but maybe less time than we think.”

Want to create mini-brains in your laboratory using Cell-Mate3D?  Contact Us to learn More!

Apoptosis, or programmed cell death, is an important cellular mechanism that is critical in development and tissue homeostasis.  Apoptosis also plays a role in cancer biology.  For example, visualizing apoptosis in response to treatments is one way to characterize potential therapies.

While 3D cell cultures are ideal for mimicking a tumor microenvironment, obtaining apoptosis data after treating the cultures can prove difficult.

To overcome this difficulty and to make apoptosis data accessible to researchers using Cell-Mate3D™ matrix, we optimized the commonly used Invitrogen™ CellEvent™ Caspase-3/7 Green Detection Reagent that enables quick and reliable imaging of apoptotic cells in culture.

Cell-Mate3D™ cultures were setup, treated, and analyzed as followed:

• AU565 human breast cancer cells (HER2-positive) were embedded into the Cell-Mate3D™ matrix.
• One sample was left untreated and an equivalent sample was treated with 100 μM Taxol for 12 days.
• Samples were stained with 15 μM CellEvent™ Caspase-3/7 reagent (three times the recommended concentration).
• Green-fluorescent apoptotic cells were clearly seen in the Taxol-treated sample by inverted confocal microscopy.
• Caspase staining appears brighter in Taxol treated cultures compared to non-Taxol treated cultures.

Figure 1. Detection of apoptotic cells in the Cell-Mate3D™ matrix using CellEvent™ Caspase-3/7 Green reagent. AU565 breast cancer cells were embedded in the Cell-Mate3D™ matrix and cultured for 12 days. Cells were untreated (LEFT) or treated (RIGHT) with 100 μM Taxol for 12 days to induce apoptosis. Cells were then incubated with the Invitrogen™ CellEvent™ Caspase-3/7 Green Detection Reagent (15 μM) for 30 min. to label apoptotic cells with green fluorescence, counterstained with the Invitrogen™ NucBlue™ Live ReadyProbe™ Reagent, and imaged using an inverted confocal microscope at 20x magnification.

Reagents Used

• Cell-Mate3D Kit – Order Now
• CellEvent™ Caspase-3/7 Reagent (ThemoFisher Scientific)
• NucBlue™ Live ReadyProbe™ Reagent (ThermoFisher Scientific)
• CellEvent, NucBlue, and ReadyProbe are trademarks of ThermoFisher Scientific

Have a question?  Call us 855 849 BRTI (2784) or email info@brtilifesciences.com

Cancerous tumors are heterogeneous tissues with a dynamic microenvironment. They exhibit an oxygen gradient with outer regions of well-oxygenated (normoxic) tissue alongside poorly-oxygenated regions experiencing hypoxia (1).

Hypoxia is essential for tumor development and many studies have shown that tumor cells in hypoxic regions distant from blood vessels show resistance to chemotherapy or radiation therapy (1).

2D cultures and the lack of an in vitro model that can recapitulate a hypoxic, 3D tumor microenvironment, are limiting to researchers who want to target cells in this context.

We show that Cell-Mate3D™ exhibits features of a tumor microenvironment by demonstrating that by day 3 in culture:

• Cells in the matrix express HIF-1α
• A hypoxic gradient within the matrix is created without the use of hypoxic chambers

Cell-Mate3D Creates a Hypoxic Gradient

The ability to mimic a physiologically relevant tumor microenvironment that contains an oxygen gradient would serve as a useful tool for cancer researchers studying tumor biology and potential treatments and therapies (4).

HIF-1α is a transcription factor that is activated when cells experience a hypoxic environment (2). When HeLa cells are embedded into the Cell-Mate3D matrix, a hypoxic gradient is formed after 3 days (Figure 1). This is demonstrated by sectioning and staining a cross section of the matrix and performing HIF-1α staining. Imaging and analysis show that the HIF-1α staining intensity is low for in the outer 400-500μm of the matrix, and that expression is increased in the center of the matrix at Day 3.

This observation indicates that culturing cells in Cell-Mate3D creates a hypoxic gradient and more closely mimics a solid tumor microenvironment compared to 2D culture.

Figure 1. Cell-Mate3D cultures were created at a density of 4 million HeLa cells per 100μL of Cell-Mate3D matrix. After one day in culture, cryosectioning and staining showed little HIF-1α expression (a,b). After three days in culture, HIF-1α staining was more intense (d,e) which is indicative of hypoxia. Relative Fluorescence Intensity (RFI) profiles of (a) and (d) show that hypoxic conditions are time dependent. After three days in culture, HIF-1α RFI (red) is elevated up to four fold compared to one day in culture (c,f). Furthermore, the intensity profile of Day 3 cultures (f) indicates the presence of an oxygen gradient. The RFI of HIF-1α begins to increase approximately 400-500μm from the edge of the matrix.

Cell stains used:

• Rabbit anti human HIF-1α (H-206) – Santa Cruz Biotech (SC-10790)
• NucBlue® Live ReadyProbes® Reagent – ThermoFisher (R37605)
• NucBlue and ReadyProbes are a registered trademark of ThermoFisher Scientific

Want to create your own hypoxic gradient without the use of expensive chambers? The Cell-Mate3D matrix is suitable for many cell types including cancer cells, fibroblasts, and stem cells.

Contact us today to schedule a free consultation on creating your own in vitro tumor microenvironment or developmental model.  Email info@brtilifesciences.com

Learn more about Cell-Mate3D

References

1. Tannock IF.  Tumor physiology and drug resistance. Cancer Metastasis Rev 20: 123–132. (2001)
2. LaGory EL, Giaccia AJ. The ever-expanding role of HIF in tumour and stromal biology. Nat Cell Biol. Apr;18(4):356-65. (2016)
3. Vaupel  P, Mayer, A. Hypoxia in cancer: significance and impact on clinical outcome. Cancer Metast. Rev. Jun;26(2):225-39. (2007)
4. Saggar JK, Yu M, Tan Q, Tannock IF (2013) The tumor microenvironment and strategies to improve drug distribution. Jun 10;3:154. (2013)

Stem cell research has been a topic of interest for a number of years. Even if you have only a vague understanding of what stem cells are, you likely know that they hold great promise for treating any number of conditions or diseases.

But there’s also a potential darker side to stem cells. Some cancer researchers have proposed a cancer stem cell hypothesis, where cancer stem cells are actually found within tumors themselves and are thought to be responsible for tumor growth, resistance, and recurrence, even after treatment. The hypothesis puts forth the idea that these cells are what makes malignant tumors so hard to treat.

However, this news isn’t all bad. Traditional cancer treatment methods like chemotherapy and radiation strive to kill all cancerous cells. Not only do these treatments often kill healthy cells in the process, but they’ve also been shown to be ineffective in treating more aggressive and advanced cancers.

The hypothesis suggests that “indiscriminate killing of cancer cells would not be as effective as selective targeting of the cells that are driving long-term growth.” If these cancer stem cells can be identified and targeted, treatments may become more effective and less exhausting for patients.

While this theory has not been completely proven, it does hold promise for the future of cancer treatments. Considering that approximately 39.6% of men and women will receive a cancer diagnosis at some point during their lives, scientific researchers understand the importance of improving existing treatments and finding new ways to study cancerous cells. At BRTI Life Sciences, our 3D cell culture systems provide yet another innovative way to more effectively study tumors and test different cancer treatments in vitro. To find out more about our tumor modeling options, please contact us today.

For many years, two-dimensional cell cultures were one of the only options researchers had to create and study cell growth and interaction. But 2D systems have definitive limitations: unlike how they behave in the human body, cells can’t grow in all directions in 2D, which makes it difficult to get realistic results. Because these cell cultures aren’t totally accurate, they don’t allow researchers to precisely predict how a given substance or medication will react in vivo.

3D tumor models, however, are much more true to life. Because 3D cell cultures mimic the tumor microenvironment more closely than 2D systems do, this realism results in much better research. These 3D tumor models accurately replicate the microenvironments of tumors found in nature like necrosis, angiogenesis, cell adhesion, and more. When researchers use 3D cell culture systems to have a more complete picture, there are more promising possibilities for the future of cancer treatments.

Since the types and behaviors of tumors are varied — there are over 120 different types of brain tumors alone — it can be a challenge for researchers to create treatments that are effective for patients across the board. However, thanks to 3D tumor models, these treatments may one day be totally personalized.

As it stands now, new cancer drugs need to be clinically tested prior to approval. It costs over $2 billion to develop just one new cancer drug, and nearly 90% of drugs that enter clinical trials fail. By using 3D cell systems, researchers can observe the exact behavior of certain cells from patient samples when given specific treatments. This gives researchers the potential to make personalized and effective cancer treatments for patients. Overall, this process would bring the cost of cancer treatments down, as time and money would not be wasted on developing treatments that don’t work in the human body.

The use of 3D models is still relatively new, but their potential is tremendous. At BRTI Life Sciences, we’ve made it simple for you to design your own 3D models for tumor research. Our Cell-Mate3D™ is a 3D cell culture matrix that is tissue-like, injectable, and chemically defined, thus offering a realistic microenvironment to be used in both in vitro and in vivo biomedical studies. To find out more about how our products can help you to conduct more accurate research, please contact us today.

Read the article in Stem Cell Translational Medicine: Rapid Induction of Cerebral Organoids From Human Induced Pluripotent Stem Cells Using a Chemically Defined Hydrogel and Defined Cell Culture Medium.

Application of Cell-MateTM 3D matrix in modeling artificial breast ductal cancer as well as control “normal” tissues…

Read more…