Stem cells are defined as cells that are capable of self-replication by mitotic cell division and subsequent differentiation into specialized cell types. Neural stem cells (NSC) are cells that are capable of self-renewal and differentiation into neurons, astrocytes and oligodendrocytes. These cells can be isolated from either fetal or adult brain and differentiated. Although, theoretically, the NSC should differentiate into all three cell types, it is often a challenge to achieve this in practice. Arriving at the correct mixture of hormones and chemicals to culture neurons, astrocytes and oligodendrocytes from NSC can be tricky. StemCell Technologies have come up with an NSC differentiation kit which contains a supplement that has a mixture of chemicals and hormones needed for optimal differentiation. This is an excellent tool to culture all 3 types of neural cells from NSC.
The Neurocult® NS-A Differentiation Kit comes with Neurocult NS-A basal medium and Neurocult NS-A differentiation supplements. The basal medium should be stored at 2-8°C and can be used with both the Neurocult NS-A proliferation and differentiation supplements. The basal medium is stable for six months and must be used with the differentiation supplements. The differentiation supplements are supplied as a frozen mixture that needs to be aliquoted and stored at -20°C. Ten ml of supplements are to be added with 90 ml of basal medium to make the complete Neurocult differentiation medium. Although the manufacturer recommends preparation of complete medium freshly before each use, our experience in the lab shows that the complete medium is stable at 4°C for about four weeks.
We have used the Neurocult NS-A Differentiation Kit in our laboratory for the last four years and have had excellent results. We were able to achieve differentiation of NSC into neurons, astrocytes and oligodendrocytes as evidenced by the expression of markers: map2a, GFAP and Gal-c, respectively. For differentiation, the NSC need to be plated on tissue culture dishes coated with either poly-l ornithine or Laminin/lysine. We have been able to generate neurons, astrocytes and oligodendrocytes using the Neurocult NS-A Differentiation kit from CD133 and nestin expressing fetal human brain derived neural stem cells .The presence of all the required hormones and chemicals as a standard mixture in the supplements makes this kit very convenient and easy to use. It also assures reproducibility. However, the drawback of using this kit is that the scientist is unaware of the exact composition of the basal medium as well as the supplements. This could pose a problem in experiments where knowledge of the medium composition is crucial.
In summary, the Neurocult NS-A differentiation kit is a very easy and convenient kit for generation of all 3 major types of neuronal cells. This would be particularly beneficial to labs that are just entering the neural stem cell arena and need an easy way to confirm the “stemness” of their cells. Additionally, the company also offers products to coat the plates and also antibodies to confirm the differentiated state of the cells.
Scientist
Children’s Research Center of Michigan
Children’s Hospital of Michigan