We thank Prof. Dr. Franklin Constantini (Columbia University, New York, NY) for providing Rosa26-YFPfl/fl mice. In adults, the blood vessels remain largely quiescent. Little is known about the mechanisms converting psychosocial stress into cellular dysfunction. H.G. Video 1 illustrates three-dimensional reconstruction of the liver vasculature of the sham-operated mouse 1 mo after irradiation-conditioned bone marrow transplantation. M. Singhal and X. Liu performed most of the experiments. How to Reach Us. Zoomed-in images are shown at the bottom. Search for other works by this author on: Interdisciplinary Research Center on Biology and Chemistry, Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences, Shanghai, China, University of Chinese Academy of Sciences, Beijing, China, State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China, German Cancer Consortium, Heidelberg, Germany, Deutsche Forschungsgemeinschaft 3 C and Fig. However, in a recent randomized, controlled phase 2 trial involving 81 patients with compensated liver cirrhosis, administration of G-CSF alone or in combination with hematopoietic stem cell (HSC) infusion failed to improve liver function or to ameliorate fibrosis (Newsome et al., 2018). We now show in zebrafish that the increase in cortisol elicited by a brief period of involuntary swimming peaks at 6 days post fertilization ... (Regierungspräsidium Karlsruhe; G-29/12). Afterward, mice were bled at regular intervals, and plasma ALT/AST levels were monitored. Augustin from Regierungspräsidium Karlsruhe, Germany). After incubation, DNA-bound c-MYC was detected with c-MYC antibody and HRP-conjugated secondary antibody by a colorimetric assay, and quantified using a standard curve. By continuing to use our website, you are agreeing to, https://creativecommons.org/licenses/by-nc-sa/4.0/, Pumping blood with self-reliance and cooperation, https://doi.org/10.1016/j.stem.2017.07.004, https://doi.org/10.1016/j.exphem.2010.04.007, https://doi.org/10.1634/stemcells.22-6-994, https://doi.org/10.1126/science.275.5302.964, https://doi.org/10.1146/annurev-cellbio-092910-154002, https://doi.org/10.1016/j.jhep.2015.02.040, https://doi.org/10.1046/j.1365-2249.1998.00726.x, https://doi.org/10.1053/j.gastro.2009.05.009, https://doi.org/10.1146/annurev-immunol-032414-112019, https://doi.org/10.1016/j.blre.2014.01.001, https://doi.org/10.1016/j.mayocp.2013.10.023, https://doi.org/10.1016/j.stem.2018.07.011, https://doi.org/10.1016/S2468-1253(17)30326-6, https://doi.org/10.1371/journal.pone.0053719, https://doi.org/10.1016/j.stem.2018.01.010, https://doi.org/10.1097/MOH.0000000000000041, A novel antiangiogenic and vascular normalization therapy targeted against human CD160 receptor, The oxysterol–CXCR2 axis plays a key role in the recruitment of tumor-promoting neutrophils, Organ-specific lymphatic vasculature: From development to pathophysiology. Ischaemia-induced AKI and renal Tx We are shaping the future We are shaping the future. In brief, mice were anaesthetized with a mixture of ketamine (100 mg/kg body weight) and xylazine (10 mg/kg body weight). 4 B). Donor chimerism in the bone marrow of recipient animals was determined after sacrificing animals. (A’ and A’’) Zoomed-in images illustrating GFP+ cells incorporated into the liver vasculature. Augustin, and J. Hu wrote the manuscript. Quantitative PCR analysis revealed higher expression of Bax, an apoptotic activator, in livers of irradiated but not partially hepatectomized mice compared with nonirradiated and partially hepatectomized mice (Fig. The generation of NDPK-B / mice … BRIEF OVERVIEW OF FOREST MANAGEMENT These two certificates have been awarded to the state operated ForstBW for their sustainable forestry management practices. After washing away unbound cells, the beads were released by trypsin/EDTA treatment. Official name: Regierungspräsidium Karlsruhe, Referat 47.2 – Baureferat Mitte Postal address: Schlossplatz 4-6 Town: Karlsruhe NUTS code: DE122 Karlsruhe Postal code: 76131 Country: Germany E-mail: abteilung4@rpk.bwl.de Telephone: +49 721926-0 Fax: +49 72193340240 Internet address(es): Main address: www.rp-karlsruhe.de. The circulatory systems of WT and CAG-GFP mice were surgically conjoined, and both mice were subjected to PHx to induce liver regeneration. 3 B). RM, GK, JK, A-EW, KT, and RP: data analysis and interpretation. To this end, YFP-labeled bone marrow cells, consisting of ∼20,000 LSK cells, were injected via tail vein into the NOD-scid gamma (NSG) mice on day 2 after PHx (Fig. 4 B). Quantitative PCR was performed with SYBR Green Master Mix (Invitrogen). The resected liver lobes were isolated and subjected to FACS analysis and cryopreservation, respectively. VECad-CreERT2xRosa26-YFPfl/fl mice with labeled ECs were intravenously injected with either PBS or 1011 viral particles of empty replication-deficient adenovirus (VB180308-1016nff; VectorBuilder Inc.). 1 G). In brief, mice were anaesthetized with 1.5% isoflurane. Brief introductions into these research areas: Photonic Materials & Devices ; Quantum Optics & Spectroscopy; Biomedical Photonics; Optical Systems ; Solar Energy . Surprisingly, though, GFP+ ECs were also detectable in livers of sham-operated mice (Fig. Publications Lines of Disconnection: The Penguin London Tube Anniversary Series ... this article analyses how Edwidge Danticat’s Claire of the Sea Light and Junot Díaz’s The Brief Wondrous Life of Oscar Wao re-write the stereotype of the paradisiacal and picturesque islands. In brief, female CB‐17 SCID mouse (7 weeks) were purchased from Charles River Laboratories (Arbresle, France) and inoculated subcutaneously with NCl … A strong increase in plasma alanine aminotransferase/aspartate aminotransferase (ALT/AST) levels was observed after CCl4 treatment, indicating liver damage (Fig. H.G. "lang":"en" FORSTBW MANAGEMENT Headquarter Ministry of Rural Affairs and Consumer Protection Kernerplatz 10 . 4, D and E). (B) The frequency of circulating LSK cells in the peripheral blood of sham-operated and PHx mice was analyzed by FACS (mean ± SD, n = 5 mice). Further, the bone marrow–derived ECs showed similar expression of cell cycle regulatory genes when compared with the resident liver ECs (Fig. 4 GWB (15 Tage nach Eingang des Nichtabhilfebescheides auf eine Rüge) wird hingewiesen. Location & Hours. "to" : ["more", "e-mail", "sms", "facebook","twitter","googleplus","linkedin"], Next, cDNA was hybridized on mouse Clariom S assay (Applied Biosystems) according to the manufacturer’s protocol. S1 C, lower panel; Fig. (E) The plot shows the count of CC3+ ECs per 1 mm2 of liver tissue (mean ± SD, n = 6 mice). Get Directions +49 721 9267404. The power of new sequencing methods now makes it possible to reinvigorate medaka genetics, in particular by establishing a near-isogenic panel derived from a single wild population. Information about authorised persons and opening procedure: Keine Bieter zugelassen. All stainings were performed in 5% FCS (in PBS) on ice for 30 min with optimal dilutions of commercially prepared antibodies. The cell suspension was further treated with 5 ml 1× ACK lysis buffer to remove erythrocytes. Kontakt für Fragen von Betroffenen im … 1, D and E), which is in line with a previous study (Langley et al., 1997), suggesting that ECs are sensitive to irradiation exposure. Dort finden Sie neben dem Antragsformular nützliche Informationen zu den Anspruchsvoraussetzungen und zur Antragstellung. All procedures were performed according to the Guide of the Care and Use of Laboratory Animals published by the National Academy of Sciences and were approved by the local authorities (Regierungspräsidium Karlsruhe AZ: 35-9185.81/G—19/12, 35-9185.81/G—229/12). Indeed, bone marrow transplant, as well as G-CSF–mediated stem cell mobilization experiments, revealed that regeneration of liver vasculature relies primarily on preexisting intact liver ECs. Approval for experiments involving animals was granted by the local Animal Welfare Committee (Regierungspraesidium Karlsruhe, Germany, reference numbers … Share. In brief, 2.5 μg of nuclear extracts isolated from cell pellets with Nuclear Extract Kit (Active Motif, Inc., Carlsbad, USA) were incubated with a synthetic oligonucleotide containing the c-MYC consensus sequence. — um die Namen der Nachunternehmer ergänztes Nachunternehmervereichnis. Log In. Cells The C26 (derived from BALB/c Colon26 murine colon adenocarcinoma) and Renca (renal cell carcinoma of BALB/c origin) cell lines were provided by the tumor-bank of the German Cancer Research Center (Heidelberg, Germany) and cultured in DMEM medium with high Glucose-4500 mg/L … (A) Experimental outline of the parabiotic model. Error bars represent mean ± SD. All animal experiments were approved by the local animal ethics committee (Regierungspräsidium Karlsruhe; Ministerium für Landwirtschaft, Umwelt und ländliche Räume, Kiel, Germany). We are most grateful for the excellent technical support of the Flow Cytometry, Genomics, Light Microscopy, and Laboratory Animal Facilities of the German Cancer Research Center and Interdisciplinary Research Center of Biology and Chemistry. 1 mo later, bone marrow chimeric mice (Fig. The mesothelium is a dynamic and specialized tissue layer that covers the somatic cavities (pleural, peritoneal, and pericardial) as well as the surface of the visceral organs such as the lung, heart, liver, bowel and tunica vaginalis testis. The distribution of UMI counts per cell … VECad-CreERT2xRosa26-YFPfl/fl mice with labeled ECs were intraperitoneally injected with either peanut oil alone or in combination with CCl4 (0.7 µl/g body weight) three times a week for a duration of 4 wk (Scholten et al., 2015). You can find all important news stories, headlines, news photos and videos about Karlsruhe. Representative images of liver sections of bone marrow chimeric mice costained with GFP, CD45, and CD31 (EC-specific surface marker). Thus, the observed induction of phosphorylated histone H2A.X and cleaved caspase-3 after irradiation, as well as the significantly reduced incorporation of GFP+ BMDMCs into the liver vasculature by a liver shield, clearly shows that whole-body irradiation had caused catastrophic damage to the liver vasculature, which led to an emergency recruitment and incorporation of bone marrow–derived cells for tissue repair and rejuvenation of organ function. The number of GFP+ ECs in PHx mice was ∼1.8× higher as compared with the sham-operated mice (Fig. We hypothesized that if BMDMCs contribute to the regenerating vasculature, then there should be GFP+ ECs incorporated into the regenerated liver in Para-WT mice after PHx. See more of Europe Direct Karlsruhe im Regierungspräsidium on Facebook. local Animal Care and Use Committee (Regierungspräsidium Karlsruhe, Germany). Approximately 5,000 LSK cells were sorted by FACSAriaIII (Becton Dickinson) and injected intravenously into nonirradiated triple transgenic Rag2−/−γc−/−KitW/Wv recipient mice. — Entsorgung = Z2 Material (Arsen belastet) = 2 000 m3. After ruling out the direct contribution of BMDMCs in two independent nonirradiation-conditioned preclinical models, we investigated the contribution of terminally differentiated resident ECs to the regeneration of liver vasculature upon PHx. Arrows indicate GFP+ ECs. Cell isolation and culture. Sections were then incubated with primary antibodies overnight at 4°C, followed by fluorophore-conjugated secondary antibodies for 2 h at room temperature. 72074 Tübingen Tel +49 7071 602-0 Mail … Total RNA was extracted from the livers of irradiated or hepatectomized mice using a Trizol isolation system (Thermo Scientific) according to the manufacturer’s instructions. PanRosaYFP mice were used as donors. The successful evaluation of bone marrow–based cellular therapies in preclinical liver regenerative models (Almeida-Porada et al., 2010; DeLeve, 2013) promoted clinical trials with either autologous bone marrow transplants or mobilization of stem/progenitor cells with the administration of G-CSF (Forbes et al., 2015). 1 A; and Video 1), suggesting that bone marrow–recruited cells had incorporated into the liver vasculature independent of the PHx-induced regenerative burst, possibly as a result of irradiation-induced vascular damage. All animal studies were performed in accordance with regulations for animal welfare and approved by the responsible agency (Regierungspraesidium Karlsruhe, Land Baden-Wuerttemberg, Karlsruhe, Germany) under the following numbers: G-264/12, G-75/13, G-29/15, G-179/16 (The last two digits represent the year of … All animal experiments were approved by the institutional and governmental Animal Care and Use Committees (IRCBC-2016-02 to J. Hu and G220/11, G213/17, and G219/17 to H.G. S3 C). S1 B) were subjected to PHx to induce liver regeneration, and the liver vasculature was analyzed 10 d after PHx. Information about authorised persons and opening procedure: Official name: Vergabekammer Baden-Württemberg, RP Karlsruhe – Referat 15, Official name: Regierungspräsidium Karlsruhe, Referat 42. After washing, protein–DNA complexes were eluted and cross-links were reversed. For Video 1, a semiautomatic surface rendering module in Imaris (Bitplane) was used to create three-dimensional volumetric surface objects corresponding either to individual cells or to the liver vascular system. Edit. (C) The plot shows the count of pH2A.X+ ECs per 1 mm2 of liver tissue (mean ± SD, n = 6 mice). Rag2−/−γc−/−KitW/Wv mice were generated as described previously (Waskow et al., 2009). 1 mo later, mice were subjected to two-thirds PHx. To prevent infection and to relieve pain, the skin wounds were treated with hydrogel containing lincomycin hydrochloride (5 mg/g) and lidocaine (4 mg/g) for 3 d. 1 mo later, blood was collected from the tail veins of all the operated mouse pairs for FACS analysis to confirm the blood chimerism. In adult mice, the liver is able to restore its original mass and structure within 10 d following PHx. Author Contributions . Future studies involving stringent and mechanistic preclinical experimental approaches will need to address these questions to possibly establish a scientific rationale for bone marrow cell–involved stem cell therapies to interfere with liver dysfunction. (E) FACS analysis of donor chimerism in CD45+ cells from peripheral blood of Rag2−/−γc−/−KitW/Wv recipients (mean ± SD, n = 4 mice).