Developing an Engineered Silk-Collagen-Based 3D Model of Polarized Neural Tissue
Published: July 31, 2024
Abstract
Source: Chwalek, K. et al., Engineered 3D Silk-collagen-based Model of Polarized Neural Tissue. J. Vis. Exp. (2015).
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 | ||
| NaCl | Sigma-Aldrich | 71382 | |
| Biopsy Punch 5 mm | World Precision Instrument | 501909 | |
| Biopsy Punch 2 mm | World Precision Instrument | 501908 | |
| Trypsin | Sigma-Aldrich | T4049-500ML | |
| DNase | Roche | 10104159001 | final concentration 0.3% |
| Soybean protein | Sigma-Aldrich | T6522-100MG | final concentration 1 mg/ml |
Citar este artículo
Developing an Engineered Silk-Collagen-Based 3D Model of Polarized Neural Tissue. J. Vis. Exp. (Pending Publication), e22396, doi: (2024).