Humanized Diabetes Mouse Model

Male NMRI-Foxn1^nu/Foxn1^nu and female SCID CB-17/Icr-Prkdc^scid/Rj mice were transplanted with human insulin-producing EndoC-βH1 cells as described^{44 (link),47 (link)}. Male NMRI-Foxn1^nu/Foxn1^nu mice were used for methodological development/validation, but all follow up experiments were performed in female SCID CB-17 mice. At the day of inoculation, 4–6 × 10⁶ EndoC-βH1 cells were seeded on a rubber toric joint (EFJM), supported in Matrigel HC (Corning) supplemented with MmVEGF-164 (1 ng/ml) (BioLegend). The cell-containing or the empty vehicle rubber rings were then inserted under the epimysium in the biceps or quadriceps femoris muscle. The mice were anesthetized with 3 % isoflurane. They received short-term analgesic (Buprenorphine 0.1 mg/kg) and long-term analgesic (3 mg/ml of Acetaminophen-supplemented water for 10 consecutive days) respectively before and after the surgery. Random glycaemia was measured weekly with an ACCU-CHEK Nano glucometer (ROCHE). Once the tumour became palpable, the mice received 20 % glucose-supplemented drinking water to counter the progressive hypoglycemia induced by the EndoC-βH1 cells. Human C-peptide was measured in plasma with a human Ultrasensitive C-peptide ELISA (Mercodia). Kelly tumour-bearing mice were generated by s.c. injections of 5 × 10⁶ cells mixed 1:1 with Matrigel (Corning) in the hind leg of female Crl:NU(NCr)-Foxn1^nu mice.