Production and Characterization of a Conditionally Immortalized Dog Beta-Cell Line from Fetal Canine Pancreas.

Production and Characterization of a Conditionally Immortalized Dog Beta-Cell Line from Fetal Canine Pancreas. Cell Transplant. 2020 Jan-Dec;29:963689720971204 Authors: Czernichow P, Reynaud K, Ravassard P Abstract Since the 1970s, rodent and human insulin-secreting pancreatic beta-cell lines have been developed and found useful for studying beta-cell biology. Surprisingly, although the dog has been widely used as a translational model for diabetes, no canine insulin-secreting beta cells have ever been produced. Here, a targeted oncogenesis protocol previously described by some of us for generating human beta cells was adapted to produce canine beta cells. Canine fetal pancreata were obtained by cesarean section between 42 and 55 days of gestation, and fragments of fetal glands were transduced with a lentiviral vector expressing SV40LT under the control of the insulin promoter. Two Lox P sites flanking the sequence allowed subsequent transgene excision by Cre recombinase expression. When grafted into SCID mice, these transduced pancreata formed insulinomas. ACT-164 is the cell line described in this report. Insulin mRNA expression and protein content were lower than reported with adult cells, but the ACT-164 cells were functional, and their insulin production in vitro increased under glucose stimulation. Transgene excision upon Cre expression arrested proliferation and enhanced insulin expression and production. When grafted in SCID ...
Source: Cell Transplantation - Category: Cytology Authors: Tags: Cell Transplant Source Type: research