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Objective To investigate the therapeutic effect of frozen-thawed murine islets which were transplanted into diabetic rats after cultured with hyperbaric oxygenated rotary cell culture system (HORCCS). Methods The purified rat islets were divided into two groups: A. In vitro experiment groups (IvEG) : The rat islets in each subgroup were cultured in HORCCS or common medium for 30 days, then evaluated for the intracellular DNA and insulin contents of islets, and the viability and insulin secreting level of islets. B. Islet transplantation experimental groups (TxEG) : The frozen-thawed islets were cultured in HORCCS or common medium for 7 days, and then transplanted into the recipients. We observed the blood glucose level (BGL) and insulin secreting level in the recipients as well as the uhrastructure change of islets in TxEG. Results The viability and insulin secreting level of islets cultured with HORCCS at 14th day were much higher than those cultured with common medium (P <0.05). The blood glucose level in recipients transplanted with islets cultured with HORCCS recovered to normal value at the 2nd week and lasted for 8 weeks. All these recipients maintained the normal glucose tolerance curve. Electronic microscopy found microchannel outlets on the surface of the frozen-thawed islets cultured with HORCCS. Conclusions Frozen-thawed islets cultured with HORCCS could establish nutrient transmission microchannels, which were not only capable of oxygen and nutrients transmission, but also improving cryopreservation solution to diffuse inside the islet cells evenly and uniformly. So this method not only lessens islet damage from cryopreservation, but also improves the effect of transplantation.
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<p><b>OBJECTIVE</b>To study the effect of xenotransplantation with pig parathyroid cells, which was prepared using cell microencapsulation technique, on the treatment of hypoparathyroidism in rats without immunosuppressor.</p><p><b>METHODS</b>Parathyroid cells were isolated from 10 healthy newborn pigs and encapsulated in alginate-polylysine-alginate (APA) membranes. Thirty-two aparathyroid Wistar rats were randomly allocated to microcapsule, non-microcapsule, empty microcapsule, and control groups. Each rat was injected intraperitoneally with encapsulated porcine parathyroid cells, free porcine parathyroid cells, empty capsules or 0.9% NaCl, respectively. Total serum calcium and parathyroid hormone levels were monitored continuously for 40 weeks. And then, the transplant beds were retrieved and subjected to morphologic and electron microscopic examination.</p><p><b>RESULTS</b>In those animals xenotransplanted with microencapsulated porcine parathyroid cells, the calcium and PTH levels were consistently within the normal range during the 40 weeks. In contrast, no therapeutic effects were observed in rats in the non-microcapsule group. Furthermore, neither empty capsules nor 0.9% NaCl were shown to have any effect on the recipient's serum calcium or PTH levels. After 40 weeks, electron microscopic examination demonstrated that the parathyroid cells within the microcapsules had survived well in vivo.</p><p><b>CONCLUSIONS</b>Xenotransplantation of microencapsulated newborn pig parathyroid cells can successfully treat hypoparathyroidism in rats without using immunosuppressive drugs. The results of this study show the possible clinical use of microencapsulated porcine parathyroid cells.</p>
Subject(s)
Animals , Rats , Animals, Newborn , Calcium , Blood , Capsules , Hypothyroidism , Therapeutics , Parathyroid Glands , Cell Biology , Transplantation , Parathyroid Hormone , Blood , Random Allocation , Rats, Wistar , Swine , Transplantation, Heterologous , MethodsABSTRACT
0.05). However, statistically significant changes of those data were found perioperatively in parathyroid gland excision group (P
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Objective To study the protective effects of ulinastatin (UTI) on canine islets isolated by automated method and to determine whether the addition of UTI during the in situ pancreatic perfusion process may improve islet recovery. Methods There was no significant difference in donor-related factors (age, gender, warm ischemia time, cold storage time, weight and pancreas weight) between the two experimental groups. Twenty donors were randomly divided into two groups. Pancreases were in situ perfused via abdominal aorta using cold hypertonic citric potassium solution (HC-A) 2 500 ml, group 1 (HC-A group, n =10). The pancreases were perfused with cold HC-A and UTI ( 10 000 U/kg), group 2 (HC-A+UTI group, n =10). Intraductal collagenase V (4℃) delivery with controlled perfusion was done, the pancreas was dissociated in system of Ricordi Chamber and their purified islets were separated with Ficoll density gradient centrifugation. The digestion time, islets remaining trapped in exocrine tissue, final islet purity, insulin and C-pep secretory activity, and islet recovery were observed. The purified islets were observed under light microscopy and electronic microscopy. Results UTI supplementation did not affect the digestion time, islets remaining trapped in exocrine tissue, final islet purify, insulin and C-pep secretory activity and their ultrastructure, and islet recovery was increased. In the HC-A+UTI group the yield of islets was [( 6.17 ? 2.86 )?10 4] IEQ, while in the HC-A group, that was [( 3.42 ? 1.47 )?10 4] IEQ ( P
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Objective To observe the influence of simulated microgravity on rat islet. Methods Isolated islet were assigned to flask-culture or bioreactor-culture. Gross structure and ultrastructure of islet were observed by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Results Islets ultrastructure on 7th day in bioreactor closely resembled fresh islets,with well-formed secretory granules and abundant mitochondria. SEM showed under microgravity islets communicating each other with cavity-like areas. Conclusions The ultrastructure of islets cultured under microgravity closely resembled fresh islets.
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Objective To detective the carcinogenesis and operation principle of cyst canceration after internal drainage(ID) operation for congenital choledocal cyst (CCC).Methods The clinical data of 25 patients with cyst carcinoma after ID operation for CCC in the past 28 years were analysed retrospectively.Results The total canceration rate after internal drainage of CCC were 30.49%(25/82); after cystoduodenostomy was 35.29%(14/51),after cystojejunostomy was 22.58%(11/31), respectively. In the 25 cases, three of them were operated with Wipple operation , 4 with tumour resection plus biliary reconstrustion operation, 4 local resection with external drainage ,14 with external drainage only. Conclusions Internal drainage of CCC should be aborted becaus of the high canceration rate after the operation.
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Objective To investigate the protective effect of losartan on acute ischemia-reperfusion(I/R)(injury) of pancreas in rats.Methods Seventy-two Wister rats were randomly divided into 3 groups:(1)(Control group);(2)Ischemia-reperfusion group:the anterior mesenteric artery and the celiac artery were(occluded) for 15 min,30min and 60min followed by 6 hours reperfusion;(3)Losartan group:losartan(40mg/kg)were administered by gavage at 12h and 1h before arterial occlusion.The pathologic changes of pancreatic tissue were observed under light microscopy;TUNEL was used to detect apoptosic of pancreatic cells;Bcl-2 expression in the pancreatic cells of rats was analyzed by immunohistochemistry technique.(Results) Losartan treatment reversed the histological abnormalities including infiltration of inflammatory cells and atrophy of acinar cells.Compared with losartan group,pancreatic tissue of I/R group exhibited increased MDA[((20.1?1.2))nmol/g and((34.9?2.6)) vs(17.9?2.1)nmol/g and(25.2?3.3)nmol/g,P
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Objective To study an improved automated method for isolation and purification of large amounts of human pancreatic islets from large mammals,and try to create conditions for preparation and isolation of large amounts of human islets.Methods An improved automated system was used to isolate and purify panreatic islets of sogs.Under the general anaesthesia(GA) condition,the pancreas of the dogs was via in situ vascular perfused using cold HC-A solution and then removed en bloc with duodenum and spleen.Then they were placed in cold UW aolutian.Intraductal collagenase-Ⅴ and pefabloc(4 ℃) was delivered with controlled perfusion.lslets were dissociated in system of Ricordi Chamber and purified islets separated with Ficoll density gradient centrifugation under controlled temperature.Digestion time,islets remaining trapped in exocrine tissue,final islet purity,insulin and C-pep secretory activity,and islet recovery were observed.The purified islets were observed under light microscope and electronic microscope after 24 h culture.Results The digestion time rate was(25.0?6.0) min,islets remaining trapped in exocrine tissue was(9.4?2.4)%,final islet purify rate was(89.7?3.5)%,islet recovery after digestion was(17.2?3.6)?104 IEQ/pancreas.Islet recovery after purification was(8.3?2.0)?104IEQ/pancreas.Insulin and C-pep secretory activity of the purified islets and their ultrastructure after 24 h culture were ideal.Conclusions Our improved automated method and facilities for isolation of canine pancreatic islets are reliable,The morphology and function of the procured islets are excellent and they can foreseeably be used for large-scale preparation of huma islets for clinical use.
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Objective To investigate the effect of Fas ligant (FasL) expression of testicular Sertoli cells on cotransplanted parathyroid cells. Methods Various numbers of rat testicular Sertoli cells with FasL expression and allogeneic parathyroid cells were cotransplanted. Allograft survival, change in cell components, apoptosis of infiltrative lymphocytes and parathyroid function were analyzed. Results The parathyroid cells when transplanted alone were all rejected, the mean survival time of allografts was (17?4)days. When parathyroid cells were cotransplanted with testicular Sertoli cells, the survival time of allografts was prolonged over 60 days. When the quantity of testicular cells was increased to 4?10~6, 6 of 8 rats maintained normal serum calcium and PTH levels throughout the follow-up period of 60 days (P