This video showcases the creation of a 3D polarized neural tissue model utilizing silk-collagen scaffolds. It details the process of seeding neuronal cells on porous silk scaffolds, incubating for cell attachment, embedding the scaffold with collagen, and underscores the significance of the supportive collagen matrix in forming a 3D polarized neural tissue model.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Silk Scaffold Preparation Preparation of silk solution from Bombyx mori cocoons Cut each cocoon into 8 equal pieces using scissors. Use about 11 cocoons for 5 g of fragmented cocoons. (15 min) Prepare 2 L of 0.02 M Na2CO3 solution and boil using a hot plate. (15 min) Weigh 5 g of fr…
Representative Results
Figure 1. PTFE mold used for silk sponge scaffold preparation. The dimensions: 10 cm diameter, 2 cm height. Figure 2. 3D brain-like tissue model. (A) Porous silk sponge scaffold. (B) Live/dead staining of neurons at day 1 upon …
Divulgaciones
The authors have nothing to disclose.
Materials
Centrifuge 5804 R
Eppendorf
poly-D-lysine
Sigma-Aldrich
P6407-5MG
final concentration 10 μg/ml, dissolved in water
Neurobasal medium
Gibco
21103049
warm up to 37 °C before use
B27 supplement 50x
Gibco
17504-044
Glutamax
Gibco
35050-061
Penicilin Streptomycin
Corning
30-002-CI
Collagen I, rat tail, 100 mg
Corning
354236
final concentration 3 mg/ml
NaOH
Sigma-Aldrich
S2770
corrosive
PBS
Sigma-Aldrich
D1283-500ML
Na2CO3
Sigma-Aldrich
223530
LiBr
Sigma-Aldrich
213225
MWCO 3500 dialysis cassettes
Thermo Fisher
66110
Heating plate
Fisher Scientific
Isotemp
Sieve
Fisher Scientific
No. 270, No. 35, No. 30
PTFE
mold made in house (Figure 1) 10 cm diameter, 2 cm height