Cultivation conditions:

Wharton’s jelly (WJ) cells was cultivated in 10 cm Petri dish and MSCs medium (DMEM/F12 with Glutamax (Gibco, Carlsbad, CA, USA)) supplemented with 10% of fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA) and 0.2% Penicillin/Streptomycin (Sigma-Aldrich, St. Louis, MO, USA) (Article Differentiation of Human Mesenchymal Stem Cells from Wharton...

2.3. Neural Induction of hWJ-MSCs

The cryopreserved hWJ-MSCs were thawed and seeded on poly-L-lysine-coated T75 flasks at a density of 2 × 103 cells/cm2 in MSCs medium (Day 0). The cells were cultured in this medium at 37 °C with 5% CO2, 21% O2, and 95% humidity for one day. After approximately 24 h when the cells were well attached to the surface, the medium was changed to NSCs induction medium (DMEM/F12 with Glutamax supplemented with 2% of FBS, 0.2% of Penicillin/Streptomycin, 1% of N2 supplement (100×) (Gibco, Carlsbad, CA, USA), and EGF 10 ng/mL (Gibco, Carlsbad, CA, USA)). The cells were maintained in this medium for three days, and then the medium was changed to NSCs proliferation medium (DMEM/F12 with Glutamax supplemented with 2% of FBS, 0.2% of Penicillin/Streptomycin, 2% of B27 supplement (50×) (Gibco, Carlsbad, CA, USA), EGF 20ng/mL (Gibco, Carlsbad, CA, USA), bFGF 20ng (BioLegend, San Diego, CA, USA)). The cells were cultured in this medium at 37 °C with 5% CO2, 21% O2, and 95% humidity for six days and the medium was changed every second day.

Dorsal root ganglion neural culture and neurite outgrowth in the presence of an MSC conditioned medium

https://www.nature.com/articles/s41598-020-61167-z

DRGs were cultured in 24-well plates on coverslips coated with poly-D-lysine (20 µg/ml) and laminin (1 µg/ml) in DMEM)

You can try using a 20 µg/ml poly-D-lysine concentration

ntroduction

Polylysine solutions are typically used to coat glassware and plasticware  for the attachment and growth of cells.

Preparation

Coating culture dishes, multiwell plates, or chamber slides

Additional information

When coating with poly-l-lysine, the efficiency of coating is relatively low when prepared with distilled water due to the acidic pH of the solution that can range between a pH of 3.3-6.6. One recommendation [1] to improve coating is to use an 8.5  pH borate buffer to dissolve the lysine, with thorough rinsing after coating. In addition to the basic techniques shown here, there are a variety of approaches recommended by vendors as shown below.

VendorRecommendationMPBio

BrainBits

Genlantis

Invitroscience

Sigma-Aldrich

Thank u all for your answers

@Alexandr Chernov,

Thanks for the Answer is it a working protocol. Some people form neuroshere and differentiate them into neurons. What you mentioned is direct protocol. Iam following indirect method. I plated 3.9*105 cells per 6 well plate and 3 day after induction I saw some aggregates, does it mean they r forming spheres or just cell artifacts ism really confused could you pls help me out. Iam attaching the pic for reference.