All that happened in the first week!
We are live.
The first week is officially in the books and we are excited to share our results! As expected, there isn’t a huge amount of information to report but we know that these initial steps are vital to the overall success of the challenge.
It is important to note that all motor neurons (healthy and ALS) used for the 12-week challenge have been derived from induced pluripotent stem cells (iPSCs). iPSCs are human somatic cells (any cell of the body except sperm and egg cells) that have been reprogrammed to a pluripotent state. They are held in high regard in regenerative medicine due to their ability to propagate indefinitely, as well as give rise to every other cell type in the body. In this case, iPSCs from healthy donors or ALS sufferers have allowed us to generate motor neurons that model healthy or ALS cell behaviour. The astrocytes, however, were isolated from healthy human fetal brain (cerebral cortex) and cryopreserved at passage 2 (P2). Astrocytes in P2 exist in a quiescent state and must undergo additional maturation for at least another 2 passages (P4) until they can be used. The process of motor neuron and astrocyte maturation has been the main focus of week 1.
Motor neuron maturation:
Both our healthy and ALS human motor neurons were thawed and seeded in a 96 well plate in the presence of motor neuron maintenance medium. The motor neurons will undergo a 10–14-day maturation process, with media changing every 2-3 days.
To be useful models of human motor neurons, cells maturing in vitro should develop mature properties that are hallmarks of motor neurons in vivo such as elaborated neuronal processes and mature electrophysiological characteristics.
Let’s look at how the maturation process is going
As we can see in each sequential update for both groups, the motor neurons are developing distinct neuronal structures such as extensive neurites and networks as well as more pronounced cell bodies (somas).
The human astrocytes were thawed in T25 flasks in astrocyte medium. They will undergo a maturation process until they reach their mature structure, with media changing every 2-3 days. Characterised by their star-like shape, astrocytes represent the most abundant cell type in the brain with a broad range of active roles including; providing physical and metabolic support for neurons, nervous system repair, modulation of synaptic transmission, and maintaining brain homeostasis.
The astrocytes are beginning to develop their classic ‘star-shape’, albeit still being in the very early stages of the maturation process.
Stay tuned at 12wdc.com, as we continue to follow the maturation of both motor neuron groups and astrocytes and prepare them for treatment.