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Archives for May 2017

Cerebral Organoids Generated using Cell-Mate3D Mentioned in Mini-Me Article

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!

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Visualizing Apoptosis in 3D Cell Cultures

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

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