Isolation and Culture of Neural Stem Cells

NSC were harvested from 11.5 day gestational age (E11.5) pan-EGFP mice (Jackson Laboratory, Bar Harbor, ME), and cultured as described (6 (link)). Fetal intestines were dissected and digested in Collagenase/ Dispase (50 μg/ml; Worthington Biochemical, Freehold, NJ) for 1h. Centrifuged tissue pellets were retrieved and cultured in Dulbecco’s Modified Eagle Medium/Ham’s Nutrient Mixture F12 (DMEM/F12; Corning, Manassas, VA) supplemented 1:1 with N₂ supplement (Gibco, Grand Island, NY), with the addition of mouse basic fibroblast growth factor (b-FGF,20 ng/ml; Sigma-Aldrich, St Louis, MO), mouse EGF (20 ng/ml; Sigma-Aldrich, St Louis, MO), 15% chicken embryo extract (Gemini, West Sacramento, CA) and 1% penicillin-streptomycin (Invitrogen, Carlsbad, CA). NSCs proliferated and grew as free-floating neurospheres, and the suspended neurospheres were collected by harvesting the culture medium. Neurospheres were confirmed to contain >95% NSC as determined by staining with the stem cell specific marker Nestin.

Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link: Access Free Full Text.

Wei J., Zhou Y, & Besner G.E. (2015). Heparin-Binding EGF-like Growth Factor and Enteric Neural Stem Cell Transplantation in the Prevention of Experimental Necrotizing Enterocolitis. Pediatric research, 78(1), 29-37.

Publication 2014

E11.5) pan-EGFP mice Collagenase/ Dispase Dulbecco’s Modified Eagle Medium/Ham’s Nutrient Mixture F12 (DMEM/F12; N2 supplement (b-FGF mouse EGF penicillin-streptomycin

Basic fibroblast growth factor Chicken Collagenase Culture medium Dispase Eagle Embryo Fetal Gestational age Intestines Mouse Nestin Nutrient Pellets Penicillin Stem cell Streptomycin Supplement Tissue

Corresponding Organization :

Other organizations : Nationwide Children's Hospital

Top 5 similar protocols

Variable analysis

independent variables

Gestational age of the pan-EGFP mice (E11.5)

dependent variables

NSC proliferation and growth as free-floating neurospheres
Percentage of NSCs in the neurospheres (determined by Nestin staining)

control variables

Collagenase/Dispase concentration (50 μg/ml)
Incubation time for tissue digestion (1h)
Culture medium composition (DMEM/F12 supplemented with N2 supplement, b-FGF, EGF, chicken embryo extract, and penicillin-streptomycin)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!