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München, Germany

Knobeloch D.,Cytonet GmbH | Ehnert S.,University of Tubingen | Schyschka L.,TU Munich | Buchler P.,TU Munich | And 8 more authors.
Methods in Molecular Biology | Year: 2012

The use of isolated human liver cells in research and development has gained increasing interest during the past years. The possible application may vary between elucidation of new biochemical pathways in liver diseases, drug development, safety issues, and new therapeutic strategies up to direct clinical translation for liver support. However, the isolation of human liver cells requires a well-developed logistic network among surgeons, biologists, and technicians to obtain a high quality of cells. Our laboratories have been involved in various applications of human liver cells and we have long-lasting experiences in human liver cell isolation and their application in R&D. We here summarize the present protocol of our laboratories for cell isolation from normal resected liver tissue, the most common tissue available. In addition, we discuss the necessary network in the clinic and quality controls to maintain human liver cells in culture and the effect of 3D extracellular matrix in cultured cells which results in preservation of hepatocyte epithelial polarity in the form of bile canaliculi and repression of epithelial to mesenchymal transitions occurring in 2D cultures. © 2012 Springer Science+Business Media, LLC.

Ochsenreither S.,Charite - Medical University of Berlin | Fusi A.,Charite - Medical University of Berlin | Wojtke S.,Charite - Medical University of Berlin | Busse A.,Charite - Medical University of Berlin | And 6 more authors.
Journal of Translational Medicine | Year: 2010

Several immunotherapeutic approaches rely on antigen-specific T-cells. Restrictions in the T-cell receptor (TCR) repertoire were reported as indicator of anti-tumor cytotoxic T-lymphocyte (CTL) response in various tumor entities. It is unclear yet whether a TCR restriction in peripheral blood mirrors the tumor compartment. We compared the expression of TCR Vβ-families for the quantification of TCR repertoire alterations in blood and tissue samples from patients with colorectal carcinoma. Blood samples from patients with colorectal carcinoma and healthy volunteers and tissue samples of normal colonic mucosa and colorectal carcinoma were analyzed. Relative Vβ-family quantification was performed based on quantitative reverse transcribed PCR. Standard deviation and average mean of the single families were determined. Two variables describing the degree of Vβ-repertoire restriction were defined. Forty-eight blood samples and 37 tissue samples were analyzed. TCR repertoire restriction was higher in blood of tumor patients than in blood of healthy controls (p < 0.05). No difference in the degree of TCR repertoire restriction was found between carcinoma and unaffected colon tissue. We found no corresponding elevated TCR families among the different compartments blood, normal colon, and carcinoma tissue of the same patient. In conclusion, we observed a repertoire restriction in peripheral blood as well as in tumor tissue of cancer patients. However, in tumor tissue, repertoire alterations were comparable to normal mucosa, suggesting compartment-specific TCR distribution rather than alterations due to tumor-T-cell interaction questioning the presence of highly restricted clonal T-cell expansions in colorectal cancer as they have been described in other, assumingly more immunogenic tumor entities. © 2010 Ochsenreither et al; licensee BioMed Central Ltd.

Karthaus M.,Klinikum Neuperlach
European Journal of Medical Research | Year: 2011

Major progress for the management of invasive aspergillosis has come from the introduction of new antifungals since the late 1990s. Although mortality of invasive aspergillosis remains as high as 30-50%. Backbone of management are prophylaxis, early diagnosis and early initiation of antifungals for reduction of invasive aspergillosis related mortality. Randomized trials have been undertaken for the prophylaxis as well as treatment of invasive aspergillosis in the last two decades. Posaconazole is recommended for prophylaxis against aspergillosis in patients treated for acute myelogenous leukemia, myelodysplastic syndrome or patients with graft versus host disease after allogeneic transplantation. Efficacy has been shown for first-line therapy of invasive aspergillosis with voriconazole and liposomal amphotericin B. Gastrointestinal resorption for the azoles posaconazole, voriconazole and itraconazole differ considerably. While oral voriconazole resportion is reduced when taken with food, posaconazole has to be taken with fatty food for optimal intestinal resorption. Beside all advances in the management of invasive aspergillosis important questions remain unresolved. This article reviews the current state of prophylaxis and treatment of invasive aspergillosis and points out clinicians unmet needs. © I. Holzapfel Publishers 2011.

Vehreschild M.J.G.T.,University of Cologne | Meissner A.M.K.,University of Cologne | Cornely O.A.,University of Cologne | Maschmeyer G.,Oncology and Palliative Care | And 8 more authors.
Haematologica | Year: 2011

Background Neutropenic patients are at risk of abdominal complications and yet the incidence and impact of these complications on patients' morbidity and mortality have not been sufficiently evaluated. We aimed to assess a clinical rule for early detection of abdominal complications leading to death or transfer to intensive care in patients with chemotherapy-associated neutropenia. Design and Methods This observational multicenter study was carried out in seven German hematology-oncology departments. For inclusion, neutropenia of at least 5 consecutive days was required. Risk factors for "transfer to intensive care" and "death" were assessed by backward-stepwise binary logistic regression analyses. Chemotherapy-associated bowel syndrome was defined as a combination of fever (T ≥37.8 °C) and abdominal pain and/or lack of bowel movement for 72 hours or more. Five hundred and twenty-one neutropenic episodes were documented in 359 patients. Results The incidence of chemotherapy-associated bowel syndrome was 126/359 (35%) in first episodes of neutropenia. Transfer to intensive care occurred in 41/359 (11%) and death occurred in 17/359 (5%) first episodes. Chemotherapy- associated bowel syndrome and duration of neutropenia were identified as risk factors for transfer to intensive care (P<0.001; OR 4.753; 95% CI 2.297-9.833, and P=0.003; OR 1.061/d; 95% CI 1.021-1.103). Chemotherapyassociated bowel syndrome and mitoxantrone administration were identified as risk factors for death (P=0.005; OR 4.611; 95% CI 1.573-13.515 and P=0.026; OR 3.628; 95% CI 1.169-11.256). Conclusions The occurrence of chemotherapy-associated bowel syndrome has a significant impact on patients' outcome. In future interventional clinical trials, this definition might be used as a selection criterion for early treatment of patients at risk of severe complications. © 2011 Ferrata Storti Foundation.

Burges A.,Ludwig Maximilians University of Munich | Shabani N.,Klinikum Neuperlach | Shabani N.,Ludwig Maximilians University of Munich | Bruning A.,Ludwig Maximilians University of Munich | Mylonas I.,Ludwig Maximilians University of Munich
Archives of Gynecology and Obstetrics | Year: 2011

Introduction Inhibins, dimeric peptide hormones composed of an alpha-subunit and one of two possible betasubunits (betaA or betaB), exhibit substantial roles in human reproduction and in endocrine-responsive tumors. However, it is still unclear if normal and cancerous cervical glandular epithelial cells as well as cervical cancer cell lines of glandular origin express the inhibin-betaA and -betaB subunits. Materials and methods Normal cervical tissue samples and a total of 10 specimens of well-differentiated adenocarcinomas of the human cervix were analyzed for inhibin-betaA and -betaB subunit expression by immunohistochemical analysis. Additionally, the cervical carcinoma cell line HeLa was analyzed by immunofluorescence and RT-PCR analysis for the expression of inhibin subunits. Results Immunolabeling of normal and malignant glandular epithelium of human cervical tissue revealed a positive staining reaction for the inhibin-betaA and -betaB subunits. Additionally, the cancer cell line HeLa synthesized both inhibin subunits. When compared to the normal cervical glandular epithelium, the expression of the inhibin beta subunits became significantly reduced in cervical adenocarcinoma tissues. Discussion In conclusion, we demonstrated a strong, though differential expression pattern of inhibin-betaA and -betaB subunits in normal and malignant glandular epithelial cells of the human uterine cervix. Although the physiological role of inhibins is still quite unclear in cervical tissue, the expression of inhibin-beta-subunits might play an important role in cervical cancer carcinogenesis, since they are significantly down-regulated during pathogenesis in cervical adenocarcinomas. © Springer-Verlag 2010.

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