Generating a Pro-Inflammatory Organ Culture Model to Simulate an Early Intervertebral Disc Disease

Samenvatting

Source: Saravi, B., et al. A Proinflammatory, Degenerative Organ Culture Model to Simulate Early-Stage Intervertebral Disc Disease. J. Vis. Exp. (2021).

This video demonstrates a technique for culturing bovine intervertebral discs in bioreactors to develop a three-dimensional organ culture model. Specific nutrient-deprived media and degenerative pressure conditions are applied to the discs, mimicking the pro-inflammatory and degenerative microenvironment observed during early-stage intervertebral disc disease. The disc heights are measured post-treatment to analyze the effects of nutrient deprivation and degenerative loading.

Protocol

All procedures involving animal samples have been reviewed and approved by the appropriate animal ethical review committee.

1. Dissection of the bovine intervertebral disc

1. Rinse the whole tail thoroughly with tap water to remove dirt and hair on the surface.
   NOTE: With intact, distal ends, a maximum of 9 intervertebral discs (IVDs) (coccygeal 1-9) per tail can be used for the experiments depending on the desired size of the IVDs. Considering the desired diameter between 15-20 mm, we used 12 bovine tails with 5 IVDs per tail for the experiments.
2. Immerse the whole tail in a box co....

Materialen

Lijst van materialen gebruikt in dit artikel

Naam	Bedrijf	Catalogusnummer	Opmerkingen
Ascorbate-2-phosphate	Sigma-Aldrich, St. Louis, USA	A8960
Band saw	Exakt Apparatebau, Norderstedt, Germany	model 30/833
Betadine	Munndipharma, Frankfurt, Germany
Corning ITS Premix	Corning Inc., New York, USA	354350
DMEM high glucose	Gibco by life technologies, Carlsbad, USA	10741574
DMEM low glucose	Gibco by life technologies, Carlsbad, USA	11564446
Ethanol for molecular biology	Sigma-Aldrich, St. Louis, USA	09-0851
Fetal Bovine Serum (FBS)	Gibco by life technologies, Carlsbad, USA	A4766801
Non-essential amino acid solution	Gibco by life technologies, Carlsbad, USA	11140050
Penicillin/Streptomycin(P/S)	gibco by life technologies, Carlsbad, USA	11548876
Phosphate Buffer Solution, tablet	Sigma-Aldrich, St. Louis, USA	P4417
Primocin	InvivoGen, Sandiego, USA	ant-pm-05
Pulsavac Jet Lavage System	Zimmer, IN,USA