Fig. 1Successful transplantation of neonatal porcine pancreatic cell clusters (NPCCs) in normal mice. NPCCs were harvested and transplanted immediately into the kidneys of normal nude mice without any manipulation. Two and eight weeks after transplantation, transplanted NPCCs were examined using immunohistochemistry. Insulin is stained red and pancytokeratin is stained green, while nuclear staining is shown in blue.
Fig. 2Effect of re-aggregation on in vivo survival of pancreatic cells after transplantation. Single-cell state or re-aggregated pancreatic cells were transplanted in the kidney capsule of normal mice. The graft was examined following hematoxylin staining or immunohistochemistry two weeks after transplantation. The black arrow point to blood vessels within the graft. Insulin is shown in red and pancytokeratin is shown in green, while nuclear staining is shown in blue. NPCCs, neonatal porcine pancreatic cell clusters.
Fig. 3Effect of repeated transfection on cell viability. Pancreatic cells were transfected repeatedly with pCEP4, pHGF, or pEBVHGF every other day up to three times. Twenty-four hours after each transfection, viable cell count was analyzed using a CCK-8 kit. Cell viabilities are expressed as percent of untransfected control cells. Data are presented as the mean ± standard error of three independent experiments.
Fig. 4Concentrations of blood glucose and body weight following transplantation of pancreatic cells transfected with pHGF or pEBVHGF. Pancreatic cells were transfected with pHGF or pEBVHGF, and re-aggregated. The re-aggregated cells were transplanted into the type 1 diabetes mouse model. Blood glucose level (A) and body weight (B) were measured every other day. Time 0 indicates the day of pancreatic cell transplantation.
Fig. 5Pancytokeratin and insulin expressions in the pancreatic cells transfected three times with pEBVHGF and re-aggregated. The cells were fixed and stained right after transplantation into the diabetes mouse model. Insulin is shown in red and pancytokeratin is shown in green, while nuclear staining is shown in blue. Imaging was performed with a confocal microscope (400×objective).
Fig. 6No expression of insulin or pancytokeratin in the transplanted pancreatic cells in a diabetes mouse model. Pancreatic cells were transfected three times with pHGF or pEBVHGF, re-aggregated, and transplanted into a type 1 diabetes mice. Survival and differentiation of the transplanted cells were examined using immunohistochemistry two and five weeks after transplantation. Insulin is stained red and pancytokeratin is stained green, while nuclear staining is shown in blue.
Fig. 7Loss of pancreatic cells transplanted into normal mice. Pancreatic cells were transfected three times with pCEP4, pHGF, or pEBVHGF and then re-aggregated. Normal nude mice were transplanted with the re-aggregated pancreatic cells in the kidney capsule. The transplanted cells were examined using immunohistochemistry one, two, and eight weeks after transplantation. Insulin is shown in red and pancytokeratin is shown in green, while nuclear staining is shown in blue.