4.1.3. Human Fetal IVD Cells

The cell cultures were derived from biopsies that were obtained in accordance with the Ethics Committee of the University Hospital of Lausanne (Ethics Protocol 51/01) and following the Federal Transplantation Program guidelines. The cell banks are managed in the Department Biobank following the regulations of the Biobank for clinical research for both the fetal and adult tissue. Specific biopsies consisting of a spinal unit of three vertebra and two discs representing tissue of 6.5 mm × 5 mm in size were obtained from fetal tissue following the voluntary interruption of pregnancy at 14–16 weeks of gestation. Biopsies were first rinsed in 1% penicillin/strepotomycin in phosphate buffer saline (PBS) and adjacent soft tissue that was delicately dissected from the main disc tissue. We thereafter prepared one IVD and two adjacent vertebrae from a fetal spine unit; primary cultures used Dulbecco's Modified Eagle's medium (DMEM) (41966-029, Gibco, Waltham, MA, USA) that was supplemented with 10% fetal bovine serum and 100 mM L-Glutamine (25030-024, Gibco). Culture conditions were at 37 ◦C under 5% CO2. One juvenile patient was also used. Cell cultures from the juvenile patient following discectomy were established as above, except type II collagenase digestion, was implemented. Cells were expanded and stored frozen in liquid nitrogen at passage 1 or 2 and the cells were thawed, expanded in monolayer at passage 3, and used for analysis following passage 4. Table 3 shows donor details.
