Striatal Neuron Medium Kit
The medium promotes differentiation, viability and maturation of iPSC-derived striatal neurons (Medium Spiny Neurons).
The differentiation protocol as described in this product’s instruction manual promotes human iPSC-derived NSCs to commit to striatal neuronal lineage expressing key markers, such as DARPP32, CALBINDIN, GABA, CTIP2 and MAP2.
The medium requires supplementing with Glial-Derived Neurotrophic Factor (GDNF; 10 ng/mL) and Brain-Derived Neurotrophic Factor (BDNF; 10 ng/mL) which are not included in the kit.
|Usage notes||The medium requires supplementing with Glial-Derived Neurotrophic Factor (GDNF) & Brain-Derived Neurotrophic Factor (BDNF)|
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ReadySet: 12 mL SureBond: 90 uL Striatal Neuron Medium: 85 mL Below we laid out the calculations: Assuming you’d like to plate the cells at our recommended density – 35,000 cells per square centimetre, you would have 43 square centimetres (or 4.5 wells of a 6-well plate) of cells. Coating: For ReadySet coating, we recommend 250 uL per square centimetre. So, you’d need 11 mL of ReadySet per vial of cells (250 uL x 43 cm2) For SureBond top coating, we recommend adding 120 uL in 6 mL PBS; and then coat 100 uL of diluted SureBond per square centimetre. So, you’d need at least 86 uL of SureBond (in 4.3 mL of PBS) per vial of cells. Medium: Assuming you’d use 6-well plates to start with your experiments, each well would require 2 mL medium. For plating 1 vial of cells (19 mL total): Spin down: 10 mL Then, 2 mL for each well x 4.5 wells = 9 mL Medium A for 1 vial of the cells (40 mL total): Day 1 – complete medium change – 2 mL for each well x 4.5 wells = 9 mL Day 4, 7, 10, 13 and 16 – 2/3 medium change (1.33 mL) – 6.7 mL total for each well x 4.5 wells = 30.2 mL Medium B for 1 vial of the cells (31 mL total): Day 19, 22, 25, 28 and 31 – 2/3 medium change – 6.7 mL x 4.5 wells = 30.2 mL Hence, you’d need 81 mL of medium per vial of cells to differentiate and mature the cells to Day 33 for assaying.