Coculture

1. Generation of mature neurons expressing specific neuronal subtype markers.

2. Differentiation can be carried out by factors secreted by mature neuronal cells without physical contact between cells.

3. The neurons generated can release neurotransmitters to the environment, demonstrating their functionality.

1. It cannot rule out the possibility of cellular fusion between MSC and mature neurons.

2. In addition to co-cultivation, the protocols require exposure to chemicals or growth factors.

3. The factors that promote differentiation are not fully identified.

4. Isolating neuronal cells is often complex and requires trained personnel to perform it.

Gene transfection

1. The tests make it possible to find determining genes for the neuronal differentiation of MSCs.

2. Most assays generate neurons with electrophysiological and phenotypic characteristics of mature neurons.

1. The transfection process is complex and often expensive.

2. Differentiation also depends on the characteristics of the cells to be differentiated from the percentage of transfection efficiency.

3. Some cells isolated from individuals with pathologies are difficult to transfect, making their application challenging to study diseases.

4. It takes exposure to other inducers.

Small molecules

1. Facilitate accurate, fast, and efficient differentiation of MSCs.

2. The effects on differentiation are specific by activating or inhibiting critical molecules in the differentiation process.

3. The effects are dose-dependent, reversible, and highly reproducible.

4. Ease of handling and storage.

5. They are less expensive than growth factors.

1. Specific cocktails may be required to generate the desired effect.

2. They have been used in recent years, but they are less known than the other differentiation protocols.