Glioblastoma Induction Using SB Mediated Transposition: A Technique for Generating Novel Mouse Model Using Transposon Mediated Integration of SB Plasmid into Neonatal Mouse Brain
Published: April 30, 2023
Abstract
Source: Calinescu, A. A. et al. Transposon Mediated Integration of Plasmid DNA into the Subventricular Zone of Neonatal Mice to Generate Novel Models of Glioblastoma. J. Vis. Exp. (2015)
This video describes an efficient technique to induce glioblastomas (GBM) using transposon DNA injected into the ventricles of neonatal mice. Cells of the subventricular zone, which take up the plasmid, transform, proliferate, and generate tumors with histopathological characteristics of human GBM.
Protocol
1. Intra-ventricular Neonatal Injections Three to four weeks prior to experiment, set up a mouse breeding cage with one male and one female mouse, and a plastic colored igloo. This provides an enriched environment and aids in the mating process. When pregnancy is confirmed, remove the male to a new cage. Eighteen days after mating, monitor cage daily for delivery. Perform intraventricular injections on postnatal day 1(P1). Preparation of the injection solution. R…
Representative Results
Figure 1: Experimental setup and guides for intra-ventricular injections in neonatal mice. (a) A stereotaxic frame (2) with micrometer dials is fitted with an automatic injector holding a 10 µl syringe (4). Inside the U frame, a neonatal adaptor frame is securely fastened (3). The control panel of the automatic injector (1) allows for precise selection of syringe, volum…
Açıklamalar
The authors have nothing to disclose.
Materials
| Stoelting's Lab Standard with Rat and Mouse Adaptors | Stoelting | 51670 | Other companies produce similar frames, any of them with small mouse adaptors are suitable |
| Quintessential Stereotaxic Injector (QSI) | Stoelting | 53311 | Injections can be made without it, ,but the automatic injector allows for increased reproducibility and convenience |
| 10 μL 700 series hand fitted MICROLITER syringe | Hamilton | Model 701 | This syringe model will deliver from 0.5 to 500 μL of solution reliably. Other syringes (for example 5 μL , Model 75) may also be used |
| 30 gauge hypodermic needle (1.25", 15^{o} bevel) | Hamilton | S/O# 197462 | This needle is ideal to pierce the skin and the skull of a neonatal mouse without the need of other invasive procedures. If it gets dull (doesn't easily enter the skin), it needs to be replaced. |
| In vivo jet PEI | Polyplus Transfection | 201-10G | Aliquot in small volumes and keep at -20ºC |
| D-Luciferin, Potassium Salt | Goldbio.com | LuckK-1g | Other sources of firefly lucifierase are just as adequate |
| Hematoxylin Solution, Harris Modified | Sigma Aldrich | HHS128-4L |
Etiketler
Bu Makaleden Alıntı Yapın
Glioblastoma Induction Using SB Mediated Transposition: A Technique for Generating Novel Mouse Model Using Transposon Mediated Integration of SB Plasmid into Neonatal Mouse Brain. J. Vis. Exp. (Pending Publication), e20692, doi: (2023).