Human hepatocyte isolation for liver cell therapy: whole marginal livers or healthy segments from splitting?
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Biomedical subjects
Publications and source records attributed to Andrea Sanna.
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Cavernous hemangioma is the most common benign liver tumor. When it becomes symptomatic, enucleation is considered the treatment of choice because of its lower morbidity compared with liver resection. Therefore, although there have been many series of laparoscopic liver resection, only a few cases of laparoscopic enucleation of liver hemangiomas have been reported. We report the case of a 36-year-old woman with a symptomatic 4-cm liver hemangioma of the left lobe who underwent laparoscopic enucleation, with complete relief of the symptoms at the 3-month follow-up. The operating time was 90 minutes. Neither liver mobilization nor ligament division was necessary. The dissection was carried out with minimal blood loss. The postoperative period was uneventful, and the patient was discharged on postoperative day 4. Laparoscopic enucleation is easy to perform in suitable lesions, and its advantage with respect to liver resection is the preservation of healthy parenchyma and liver ligaments. No outflow occlusion is needed, and the intraoperative bleeding is easily controlled. Technical aspects of laparoscopic enucleation are described and a review of the literature is also provided.
Hepatocytes transplantation is viewed as a possible alternative or as a bridge therapy to liver transplantation for patients affected by acute or chronic liver disorders. Very few data regarding complications of hepatocytes transplantation is available from the literature. Herein we report for the first time a case of portal vein thrombosis after intraportal hepatocytes transplantation in a liver transplant recipient. A patient affected by acute graft dysfunction, not eligible for retransplantation, underwent intraportal infusion of 2 billion viable cryopreserved ABO identical human allogenic hepatocytes over a period of 5 h. Hepatocytes were transplanted at a concentration of 14 million/ml for a total infused volume of 280 ml. Doppler portal vein ultrasound and intraportal pressure were monitored during cell infusion. The procedure was complicated, 8 h after termination, by the development of portal vein thrombosis with liver failure and death of the patient. Autopsy showed occlusive thrombosis of the intrahepatic portal vein branches; cells or large aggregates of epithelial elements (polyclonal CEA positive), suggestive for transplanted hepatocytes, were co-localized inside the thrombus.
Cryopreserved human hepatocytes could be the best type of cells to be used in a bioartificial liver (BAL) device due to reduced biosafety and biocompatibility risks. Banking of primary human hepatocytes, obtained from livers unwanted for transplantation at harvesting, could be used as a source of human liver cells for BAL treatment. We describe herein for the first time the case of a patient affected by fulminant hepatic failure (FHF) due to acute HBV infection that was successfully bridged to emergency liver transplantation by BAL treatment using cryopreserved primary human hepatocytes. The use of cryopreserved primary human hepatocytes as the biological part of the BAL device has never been described before and might be considered as a possible alternative to xenogenic material or human tumoral cell lines due to reduced biosafety and biocompatibility risks.
The offer of liver transplantation to many patients affected by liver failure is limited by organ shortage. Clinical application of human-based liver cell therapies, such as bioartificial liver and hepatocyte transplantation, might support liver transplantation, allowing more patients to be treated and decreasing mortality in the waiting list. The development of a standardized method of hepatocyte isolation is a mainstay for large-scale application of liver cell therapy. The aim of this study is to analyze retrospectively a 2-year experience of human hepatocyte isolation from livers rejected from transplantation at organ harvesting, performed on a national basis in Italy. All the livers judged unsuitable for transplantation were considered for hepatocyte isolation. Macrosteatosis greater than 60% was the most common reason of refusal, followed by nonviral cirrhosis. Fifty-four organs were used. Human hepatocyte isolation resulted in more that 7 million liver cells/g of tissue digested with 73% +/- 14% viability. Steatotic organs gave better results in terms of cell yield than cirrhotic livers. Isolated hepatocytes were able to perform specific liver functions, and evidence of factor IX and albumin messenger RNA (mRNA) production was reported when cells were plated in culture. Modifications of the traditional method of hepatocyte isolation, aimed at reducing ischemia-reperfusion damage and improving post-isolation cell conditions, showed improvements in post-isolation viability. In conclusion, we show that it is possible to use the vast majority of livers not suitable for transplantation on a national basis for human hepatocyte isolation, obtaining a large amount of viable functioning human hepatocytes that might be used for cell transplantation and therapy.
Mobile devices such as Personal Digital Assistants, Tablet PCs, and cellular phones have greatly enhanced user capability to connect to remote resources. Although a large set of applications are now available bridging the gap between desktop and mobile devices, visualization of complex 3D models is still a task hard to accomplish without specialized hardware. This paper proposes a system where a cluster of PCs, equipped with accelerated graphics cards managed by the Chromium software, is able to handle remote visualization sessions based on MPEG video streaming involving complex 3D models. The proposed framework allows mobile devices such as smart phones, Personal Digital Assistants (PDAs), and Tablet PCs to visualize objects consisting of millions of textured polygons and voxels at a frame rate of 30 fps or more depending on hardware resources at the server side and on multimedia capabilities at the client side. The server is able to concurrently manage multiple clients computing a video stream for each one; resolution and quality of each stream is tailored according to screen resolution and bandwidth of the client. The paper investigates in depth issues related to latency time, bit rate and quality of the generated stream, screen resolutions, as well as frames per second displayed.
The Roux-en-Y gastric bypass (RYGBP) and the biliopancreatic diversion (BPD) induce long-term control of type 2 diabetes in morbidly obese individuals. The reasons for such an effect on glycemic metabolism are thought to be secondary to reduced food intake, weight loss and modifications of the enteroinsular axis which is impaired in type 2 diabetic patients. Both GLP-1 and GIP have an impaired secretin effect in type 2 diabetics, and surgery can restore this function. GIP is a peptide secreted by the duodenal K-cells in response to ingested fat and carbohydrate. In obese type 2 diabetes patients, its receptor on beta-cells is down-regulated. GLP-1 is a peptide secreted by the gut L-cells, and, in type 2 diabetes, its secretion is impaired. Both RYGBP and BPD provide durable GLP-1 delivery, both during fasting and after meal ingestion, inducing L-cell stimulation by early arrival of nutrients in the distal ileum. The secretion of GLP-1 influences glucose metabolism by inhibiting glucagon secretion, stimulating insulin secretion, delaying gastric emptying and stimulating glycogenogenesis. In conclusion, the early arrival of a meal in the terminal ileum seems to be the common feature of both operations that leads to an improvement in glycemic metabolism and to resolution of type 2 diabetes.
The discovery of c-kit gene mutations and the positivity of its transcription products in gastrointestinal stromal tumours (GISTs) suggest a possible origin from Cajal interstitial cells. The study population consisted of 12 patients with GIST, with a mean age of 67.6 years. Preoperative biopsy was performed in 4 cases and in only 1 case did it prove correct. Mesenchymal tumours were regarded as myogenic when they were positive for desmin, and as neurogenic when they were positive for S-100 protein or specific neural enolase (SNE). Seven out of 12 patients underwent simple tumour excision, while in 2 cases ileal resection was performed; gastric resection, total gastrectomy with D2 lymphadenectomy and left colectomy were carried out in one case each. There were two deaths, both unrelated to the primitive diagnosis. Immunohistochemical studies were positive for CD34 in 58% of the cases, and for CD117 in 83%. The mitotic count was higher than 5/10 HPF in 3 cases. The mean survival was 57 months. The overall survival rate was 66%. We found that good tumour differentiation, small size and a low mitotic index correlate with benign behaviour and a better prognosis. Positivity for CD117, evaluated in all malignant lesions, was slightly lower (83% vs 89%) as compared to the data reported in the literature. Tumours with a high mitotic index (> 5 mitoses/10 HPF) and measuring more than 5 cm in diameter are to be considered malignant.