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Xenøgeneic engraftment of porcine fetal lateral ganglionic eminence fells into the rat hippocampus: A potential therapy for epilepsy
Cell Transplantation, Vol. 5, No. 5S-2, pp. 31-36, 1996 Copyright © 1996 Elsevier Science Inc. Printed in the USA. All rights reserved 0963-6897/96 $15.00 + .00 EI,SEV1ER
PII S0963-6897(96)00163-7
Engraftment and Primary Non-Function Category 4
4.01 BOVINE TO PORCINE INTRATHYMIC ISLET XENO-
TRANSPLANTATION Coppdli A, Giannarelli R, ]3el Guerra S, Carmellini M, Solari R, Calabrese M, Tellini C, Lupi R, Marchetti P, Navalesi R; Pisa ITALY Intrathymic islet transplantation allows survival of auto- and allo-islet grafts in rodent, dog and pig experimental models. In addition, the thymus is a suitable site for islet survival in concordant xeno-transplantation rodent models (rat-to-mouse). In this study we prepared pure bovine islets, of woven in-vitro and in-vivo viability, and cultured them at 37 °C in medium M199, with 10% bovine serum and antibiotics for 15 to 21 days. Then, approximately I0,000 islets were transplantanted into the thymus of eight 3-too old, non-immunosuppressedpigs, under general anesthesia. The surgical procedure was simple, safe and effective. The glands were retrieved after 3 (2 animals), 7 (3 animals) and 14 (3 animals) days from implantation, and processed for light microscopy (hematoxylin-cosin staining, immunocytochemisU'y). In 2 pigs of the 14 day study, intravenous glucose was administered at 3, 7 and 14 days afar islet transplantation, and bovine insulin measured by RIA following HPLC separation from porcine insulin. Well preserved islets were shown both at 3 and 7 days from transplantation, and insulin containing cells were evidentiated by immunocytochemical staining. No apparent modification of the islet surrounding thymus was observed. Bovine insulin was found in the plasma of transplanted pigs, which increased upon glucose stimulation, up to 14 days from implantation. These results demonstrate that bovine islets can survive for a few days in the thymus of non-immunosnppressed pigs, with avoidance of hyperacute rejection. Further studies ure ongoing to evaluate how long the islets can function in this discordant xenograft model.
4.02 XEN(~GENEIC ENGRAFTMENT OF PORCINE FETAL
LATERAL GANGLIONIC EMINENCE CELLS INTO THE RAT HIPPOCAMPUS: A POTENTIAL THERAPY FOR EPILEPSY Jacoby DB, Lindberg C, Ratliff J, Beaulieu L, Bousquet J, Dinsmore J; Charlestown USA Considerable research has demonstrated that intracerebral engraftment of fetal neural cells can potentiate the severity of epileptic seizures in animal models. Noradrenergic locus ceruleus neurons have been used to reinnervate the hippocampus and normalize basal and seizure-induce noradrenaline levels in the host brain. However, while some reports are promising, others have demonstrated only mild anticonvulsant effects. Recently, the inhibitory neurotransmitter gamma aminobutyric acid (GABA) has been shown to suppress seizures, thus raising the possibility that GABAergic tissues such as fetal lateral ganglionic eminence (LGE) cells may provide a regulated source of GABA. Porcine fetal LGE cells have previously been shown to survive and integrate in the adult rat striatom, but work by others has shown poor survival of fetal striatal cells outside of the striatum. We now adch'esswhether ectopically placed porcine LGE cells can survive and integrate in the hippocampus, a central regulator of epileptic activity. Our preliminary data demonstrates that fetal pig striatal grafts survive long term within the hippocampus.
PII S0963-6897(96)00163-7
Engraftment and Primary Non-Function Category 4
4.01 BOVINE TO PORCINE INTRATHYMIC ISLET XENO-
TRANSPLANTATION Coppdli A, Giannarelli R, ]3el Guerra S, Carmellini M, Solari R, Calabrese M, Tellini C, Lupi R, Marchetti P, Navalesi R; Pisa ITALY Intrathymic islet transplantation allows survival of auto- and allo-islet grafts in rodent, dog and pig experimental models. In addition, the thymus is a suitable site for islet survival in concordant xeno-transplantation rodent models (rat-to-mouse). In this study we prepared pure bovine islets, of woven in-vitro and in-vivo viability, and cultured them at 37 °C in medium M199, with 10% bovine serum and antibiotics for 15 to 21 days. Then, approximately I0,000 islets were transplantanted into the thymus of eight 3-too old, non-immunosuppressedpigs, under general anesthesia. The surgical procedure was simple, safe and effective. The glands were retrieved after 3 (2 animals), 7 (3 animals) and 14 (3 animals) days from implantation, and processed for light microscopy (hematoxylin-cosin staining, immunocytochemisU'y). In 2 pigs of the 14 day study, intravenous glucose was administered at 3, 7 and 14 days afar islet transplantation, and bovine insulin measured by RIA following HPLC separation from porcine insulin. Well preserved islets were shown both at 3 and 7 days from transplantation, and insulin containing cells were evidentiated by immunocytochemical staining. No apparent modification of the islet surrounding thymus was observed. Bovine insulin was found in the plasma of transplanted pigs, which increased upon glucose stimulation, up to 14 days from implantation. These results demonstrate that bovine islets can survive for a few days in the thymus of non-immunosnppressed pigs, with avoidance of hyperacute rejection. Further studies ure ongoing to evaluate how long the islets can function in this discordant xenograft model.
4.02 XEN(~GENEIC ENGRAFTMENT OF PORCINE FETAL
LATERAL GANGLIONIC EMINENCE CELLS INTO THE RAT HIPPOCAMPUS: A POTENTIAL THERAPY FOR EPILEPSY Jacoby DB, Lindberg C, Ratliff J, Beaulieu L, Bousquet J, Dinsmore J; Charlestown USA Considerable research has demonstrated that intracerebral engraftment of fetal neural cells can potentiate the severity of epileptic seizures in animal models. Noradrenergic locus ceruleus neurons have been used to reinnervate the hippocampus and normalize basal and seizure-induce noradrenaline levels in the host brain. However, while some reports are promising, others have demonstrated only mild anticonvulsant effects. Recently, the inhibitory neurotransmitter gamma aminobutyric acid (GABA) has been shown to suppress seizures, thus raising the possibility that GABAergic tissues such as fetal lateral ganglionic eminence (LGE) cells may provide a regulated source of GABA. Porcine fetal LGE cells have previously been shown to survive and integrate in the adult rat striatom, but work by others has shown poor survival of fetal striatal cells outside of the striatum. We now adch'esswhether ectopically placed porcine LGE cells can survive and integrate in the hippocampus, a central regulator of epileptic activity. Our preliminary data demonstrates that fetal pig striatal grafts survive long term within the hippocampus.