Papers by Werner Lindenmaier
Genetic Engineering Techniques: Recent Developments, 1982
BIOspektrum, 2011
Zusammenfassung Einweg-Beutelsysteme sind die Basis für die sichere Herstellung therapeutischer ... more Zusammenfassung Einweg-Beutelsysteme sind die Basis für die sichere Herstellung therapeutischer Zellen. Durch Atmosphärendruck-Plasmaverfahren modifizierte Oberflächen ermöglichen Zelladhäsion und stellen Gruppen für die Kopplung biologisch aktiver Substanzen bereit.
Proceedings of the National Academy of Sciences, 1984
A human cosmid library was constructed and probed with a human alpha interferon (IFN-alpha) cDNA ... more A human cosmid library was constructed and probed with a human alpha interferon (IFN-alpha) cDNA clone. One clone giving a strong hybridizing signal was isolated and characterized. The cosmid DNA insert represents a section of the human genome containing three regions of IFN-alpha-like sequences. The DNA was characterized with restriction endonuclease mapping, thereby allowing comparison to similar linkage groups reported recently and determination of homologous regions on the known physical map. The three IFN-alpha-like sequences were analyzed by a partial sequence analysis. Mapping and sequence data establish this section as a not-yet-described cluster of IFN-alpha sequences in the human genome; however, a part of the section matches to some degree to a previously described genomic region. The region described here could represent genetic polymorphism or a duplicated segment.
MGG Molecular & General Genetics, 1978
In vitro recombination techniques were used to construct hybrid plasmids between pBR 322 and diff... more In vitro recombination techniques were used to construct hybrid plasmids between pBR 322 and different DNA fragments derived from pML 21 and the E. coli chromosome. Some of the resulting hybrid plasmids express the tetracycline resistance gene of pBR 322, depending on the DNA fragment which has been ligated into the HindIII-site ofpBR 322.
STEM CELLS, 2010
Ligament-to-bone and tendon-to-bone interfaces (entheses, osteotendinous junctions [OTJs]) serve ... more Ligament-to-bone and tendon-to-bone interfaces (entheses, osteotendinous junctions [OTJs]) serve to dissipate stress between soft tissue and bone. Surgical reconstruction of these interfaces is an issue of considerable importance as they are prone to injury and the integration of bone and tendon/ligament is in general not satisfactory. We report here the stem cell-dependent spontaneous formation of fibrocartilaginous and fibrous entheses in heterotopic locations of the mouse if progenitors possess a tenogenic and osteo-/chondrogenic capacity. This study followed the hypothesis that enhanced Bone Morphogenetic Protein (BMP)-signaling in adult mesenchymal stem cells that are induced for tendon formation may overcome the tendon-inherent interference with bone formation and may thus allow the stem cell-dependent formation of tendon-bone interfaces. The tenogenic and osteo-/chondrogenic competence was mediated by the adeno- and/or lentiviral expression of the biologically active Smad8 signaling mediator (Smad8ca) and of Bone Morphogenetic Protein 2 (BMP2). Modified mesenchymal progenitors were implanted in subcutaneous or intramuscular sites of the mouse. The stem cell-dependent enthesis formation was characterized histologically by immunohistological approaches and by in situ hybridization. Transplantation of modified murine stem cells resulted in the formation of tendinous and osseous structures exhibiting fibrocartilage-type OTJs, while, in contrast, the viral modification of primary human bone marrow-derived mesenchymal stromal/stem cells showed evidence of fibrous tendon-bone interface formation. Moreover, it could be demonstrated that Smad8ca expression alone was sufficient for the formation of tendon/ligament-like structures. These findings may contribute to the establishment of stem cell-dependent regenerative therapies involving tendon/ligaments and to the improvement of the insertion of tendon grafts at bony attachment sites, eventually.
International Journal of Oral and Maxillofacial Surgery, 1999
Bioresorbable materials are of growing importance in the reconstruction of osseous defects in ora... more Bioresorbable materials are of growing importance in the reconstruction of osseous defects in oral and maxillofacial surgery. Their use instead of autogenous bone grafts reduces postoperative morbidity significantly.
Zhonghua shao shang za zhi = Zhonghua shaoshang zazhi = Chinese journal of burns, 2005
To investigate the influence of vascular endothelial growth factor (VEGF) gene modification on sk... more To investigate the influence of vascular endothelial growth factor (VEGF) gene modification on skin substitute grafted on nude mice. Human fibroblasts were transfected with VEGF adenovirus vector. Then the genetic modified fibroblasts were seeded on patches of Integra artificial skin. Twenty-four hours later, the Integra patches were grafted onto full-thickness skin defects on nude mice. Seventy-two nude mice were divided into experiment (n = 18, E, with fibroblasts seeded on Integra which were transfected by adenovirus containing VEGF in advance), GFP control (n = 18, the fibroblasts were transfected with adenovirus containing labelled GFP segment as same as that in E group, but containing no VEGF gene), Fb control (n = 18, without gene transfection), and control (n = 18, no fibroblast was seeded on Integra) groups. The survival rate, the revascularization process and the histological changes in the grafts in gene modified group (experimental group) and control groups were observed...
Langenbeck's Archives of Surgery, 2011
Background With the development of cell-based gene transfer techniques, genetically modified huma... more Background With the development of cell-based gene transfer techniques, genetically modified human keratinocytes (Kc) and fibroblasts (Fb) have been proven to be a better choice in wound repair. Methods This study was designed to construct in one step a gene-modified artificial skin by a genetically engineered Kc expressing PDGF-BB and Fb expressing VEGF 165 and bFGF. The wound healing effect in a full-thickness wound model was then observed. Unmodified artificial skin served as control. On the post-operative days 7, 14, and 21, residual wound area was calculated and skin wound tissues were subjected to biopsy for further investigation. Results Compared with unmodified artificial skin, genemodified artificial skin resulted in a reduced wound contraction and a well-organized human epidermis and better formed dermis.
Journal of Medical Virology, 1997
Although a T-cell response in human cytomegalovirus (HCMV)-immune individuals exists against the ... more Although a T-cell response in human cytomegalovirus (HCMV)-immune individuals exists against the most abundantly expressed protein pp65 of the virus matrix, less is known about the determinants that evoke this response. The aim of the study was to identify regions within HCMV pp65 (ppUL83) that contain sequences for the cellular immune response by the use of three recombinant overlapping beta-galactosidase pp65 fusion proteins (C74, C35, and C47), covering the C-terminal 265 amino acids of the entire pp65 sequence. Two T-cell epitope determinants were recognized by human lymphocytes of healthy, HCMV-seropositive, human leukocyte antigen (HLA)-typed individuals. One T-cell determinant (amino acids [aa] 303-388) was localized in the mid-region of the entire pp65 sequence and a second T-cell determinant (aa 477-561) within the C-terminal region. By fine mapping with synthetic hexadecamer peptides three T-cell epitopes were identified within these two regions: P10-I (aa 361-376) in the mid-region, P3-II (aa 485-499), and P6-II (aa 509-524) in the C-terminal region. Inhibition studies with monoclonal antibodies to HLA class I or class II revealed a class II restricted response to peptides P10-I or P6-II, respectively. P10-I responders shared the HLA-DR11 allele and P6-II responders the -DR3 allele. Therefore, these T-cell epitopes of HCMV pp65 might be presented in association with particular HLA class II alleles.
Contributions to microbiology and immunology, 1979
Nar, 1984
A phage library and two cosmid libraries were screened for human VK genes. Two recombinant phage ... more A phage library and two cosmid libraries were screened for human VK genes. Two recombinant phage and four cosmid clones were analysed in detail by restriction mapping and sequencing. Each one contained a single VKI sequence. Two of these six sequences are potentially functional VK genes and four are pseudogenes. Two pseudogenes derived from different genomic DNAs are highly homologous and are therefore either allelic variants or the products of a recent duplication event. Comparisons of our sequences with all fully determined human VKI amino acid and DNA sequences reveal identical segments which at first sight appear like minigenes. But these segments do not coincide with the subregions and some of the segments include both, framework and complementarity determining regions (FR, CDR, ref. 2). The findings may be explained by an evolutionary model generating composite genes by gene conversion and selection.
Nucleic Acids Research, 1984
A phage library and two cosmid libraries were screened for human VK genes. Two recombinant phage ... more A phage library and two cosmid libraries were screened for human VK genes. Two recombinant phage and four cosmid clones were analysed in detail by restriction mapping and sequencing. Each one contained a single VKI sequence. Two of these six sequences are potentially functional VK genes and four are pseudogenes. Two pseudogenes derived from different genomic DNAs are highly homologous and are therefore either allelic variants or the products of a recent duplication event. Comparisons of our sequences with all fully determined human VKI amino acid and DNA sequences reveal identical segments which at first sight appear like minigenes. But these segments do not coincide with the subregions and some of the segments include both, framework and complementarity determining regions (FR, CDR, ref. 2). The findings may be explained by an evolutionary model generating composite genes by gene conversion and selection.
Transfusion, Apr 1, 2010
BACKGROUND: Dendritic cells (DCs) are applied worldwide in several clinical studies of immune the... more BACKGROUND: Dendritic cells (DCs) are applied worldwide in several clinical studies of immune therapy of malignancies, autoimmune diseases, and transplantations. Most legislative bodies are demanding high standards for cultivation and transduction of cells. Closed-cell cultivating systems like cell culture bags would simplify and greatly improve the ability to reach these cultivation standards. We investigated if a new polyolefin cell culture bag enables maturation and adenoviral modification of human DCs in a closed system and compare the results with standard polystyrene flasks. STUDY DESIGN AND METHODS: Mononuclear cells were isolated from HLA-A*0201-positive blood donors by leukapheresis. A commercially available separation system (CliniMACS, Miltenyi Biotec) was used to isolate monocytes by positive selection using CD14-specific immunomagnetic beads. The essentially homogenous starting cell population was cultivated in the presence of granulocyte-macrophage-colony-stimulating factor and interleukin-4 in a closed-bag system in parallel to the standard flask cultivation system. Genetic modification was performed on Day 4. After induction of maturation on Day 5, mature DCs could be harvested and cryopreserved on Day 7. During the cultivation period comparative quality control was performed using flow cytometry, gene expression profiling, and functional assays. RESULTS: Both flasks and bags generated mature genetically modified DCs in similar yields. Surface membrane markers, expression profiles, and functional testing results were comparable. The use of a closedbag system facilitated clinical applicability of genetically modified DCs. CONCLUSIONS: The polyolefin bag-based culture system yields DCs qualitatively and quantitatively comparable to the standard flask preparation. All steps including cryopreservation can be performed in a closed system facilitating standardized, safe, and reproducible preparation of therapeutic cells. ABBREVIATIONS: BrdU = bromodeoxyuridine; DC(s) = dendritic cell(s); eGFP = enhanced green fluorescent protein; GFP = green fluorescent protein; moi = multiplicity of infection. From the
Animal Cell Technology: From Target to Market, 2001
Developmental dynamics : an official publication of the American Association of Anatomists, 2006
Three-dimensional cardiomyocyte cultures offer new possibilities for the analysis of cardiac cell... more Three-dimensional cardiomyocyte cultures offer new possibilities for the analysis of cardiac cell differentiation, spatial cellular arrangement, and time-specific gene expression in a tissue-like environment. We present a new method for generating homogenous and robust cardiomyocyte tissue cultures with good long-term viability. Ventricular heart cells prepared from fetal rats at embryonic day 13 were cultured in a scaffold-free two-step process. To optimize the cell culture model, several digestion protocols and culture conditions were tested. After digestion of fetal cardiac ventricles, the resultant cell suspension of isolated cardiocytes was shaken to initialize cell aggregate formation. In the second step, these three-dimensional cell aggregates were transferred onto a microporous membrane to allow further microstructure formation. Autonomously beating cultures possessed more than 25 cell layers and a homogenous distribution of cardiomyocytes without central necrosis after 8 we...
Nucleic acids research, Jan 25, 1982
Successful shuttling of cloned DNA in eukaryotic cells should allow isolation of expressed genes.... more Successful shuttling of cloned DNA in eukaryotic cells should allow isolation of expressed genes. We tested the utility of cosmids for moving DNA into and out of eukaryotic cells. The unique cleavage of DNA at the cos site by the terminase function of lambda was exploited to maintain the linkage between the vector and inserted gene sequences, a prerequisite for successful rescue of the transforming DNA from high molecular weight DNA of the eukaryotic transformant. A cosmid recombinant containing the HSV thymidine kinase gene and a lambda recombinant containing the chicken thymidine kinase gene were used to test the feasability of this method. It was found that these recombinants can be rescued with high efficiency from DNA of HAT-resistant cells.
Uploads
Papers by Werner Lindenmaier