Inthe Protocol of neuronal cells in one step

Inthe Protocol of neuronal cells in one step

Macro-spectrum: Can MSCs also induce neuronal adult cells? Protocol Our technology is sorted out, waiting for you to read!

First on the medium:

The operating steps of inducing neuronal cells are as follows:

1. Prepare MSCM cell culture medium and cell coated solution (Hongda Biology, No number: SD0044)

MSCM growth media were used to inoculate MSCs in liquid-coated culture bottles at a density of 4000 cells / cm2.

2 Rechanged every 1-2 days and the cells were grown to 80% saturation

Now we are ready to induce!

一、Induced the differentiation of MSC

Cultures were induced using stem cell neural differentiation medium, while one sample was cultured with MSCM growth medium as a negative control.

In the case of 6-well plates, induction started immediately after cells reached more than 80% of confluence,

1. The required induced cells were washed twice with D-PBS to remove the growth medium;

2. Add about 1ml of stem cell neural differentiation medium per well and then incubated in a 37 incubator

3. Induced culture for at least 3 days, during which the medium was changed every 48 hours. Note that significant morphological changes in the cells may be observed by 1 day after induction.

Small matters to be noted: please drop slowly along the hole wall; if the cell background is clean, please be careful to rinse the cells with PBS during the cell induction, which is easy to cause cell shedding;

Give students pictures of our induction: (induction of BMSCs in Rmsc-Bm / SD rats)

Day 1: confluence before induction of Rmsc-bm cells

Day 3: the third day of RMSC-bm induction

Day 8: Day 8 of the RMSC-bm induction

Harvest and validation of the cells

Neuron validation is the simplest way to verify the shock, but many laboratory no electric shock material, we come up with a good way, buy a lighter, take down the ignition battery, put cell culture hole gently, neuronal cells immediately show its unique contraction reaction, the operation should pay attention to press the fire, if the current is too high, cells will be, this method for just validation, do not need to send pepar students, pepar classmates still need to step by step;

MSC-derived nerve cells can selectively further verify the characteristics of cells according to the "detection of nerve cell markers"; the induced MSC cells have the main NeuN, Nestin and other neurons; immunofluorescence staining is required here, interested students can review the detailed introduction of cell immunofluorescence staining operations, in addition to WB verification.