Solid tissues can be manipulated <i>ex vivo</i> and used as vehicles for gene therapy

Hasson, Eilat; Slovatizky, Y; Shimoni, Yael; Falk, Haya; Panet, Amos; Mitrani, Eduardo

doi:10.1002/jgm.740

Cited by 30 publications

(30 citation statements)

References 41 publications

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“…Tissues, delivered within 4 h after surgery, were kept on ice until sectioning. For the preparation of decidual organ cultures, tissues were washed with phosphate-buffered saline (PBS), cut by a microtome (Tissue Sectioner, model TC-2; Sorvall Corp.) into thin slices (thickness, 250 m) encompassing ϳ10 cell layers (25), and incubated in DMEM with 25% Ham's F-12 medium, 10% fetal bovine serum, 5 mM HEPES, 2 mM glutamine, 100 IU/ml For infection of decidual organ cultures, the tissues were placed in 48-well plates (ϳ5 slices/well to maintain optimal viability) immediately after the sectioning and were inoculated with the virus (10 4 PFU/well unless otherwise stated) for 12 h to allow effective viral adsorption. Following viral adsorption, the cultures were washed extensively and were further incubated for the duration of the experiment, with replacement of the culture medium every 2 to 3 days.…”

Section: Cells and Virusesmentioning

confidence: 99%

Modeling of Human Cytomegalovirus Maternal-Fetal Transmission in a Novel Decidual Organ Culture

Weisblum

Panet

Zakay‐Rones

et al. 2011

J Virol

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Human cytomegalovirus (HCMV) is the leading cause of congenital infection, associated with severe birth defects and intrauterine growth retardation. The mechanism of HCMV transmission via the maternal-fetal interface is largely unknown, and there are no animal models for HCMV. The initial stages of infection are believed to occur in the maternal decidua. Here we employed a novel decidual organ culture, using both clinically derived and laboratory-derived viral strains, for the ex vivo modeling of HCMV transmission in the maternal-fetal interface. Viral spread in the tissue was demonstrated by the progression of infected-cell foci, with a 1.3-to 2-log increase in HCMV DNA and RNA levels between days 2 and 9 postinfection, the expression of immediate-early and late proteins, the appearance of typical histopathological features of natural infection, and dose-dependent inhibition of infection by ganciclovir and acyclovir. HCMV infected a wide range of cells in the decidua, including invasive cytotrophoblasts, macrophages, and endothelial, decidual, and dendritic cells. Cell-to-cell viral spread was revealed by focal extension of infected-cell clusters, inability to recover infectious extracellular virus, and high relative proportions (88 to 93%) of cell-associated viral DNA. Intriguingly, neutralizing HCMV hyperimmune globulins exhibited inhibitory activity against viral spread in the decidua even when added at 24 h postinfection-providing a mechanistic basis for their clinical use in prenatal prevention. The ex vivo-infected decidual cultures offer unique insight into patterns of viral tropism and spread, defining initial stages of congenital HCMV transmission, and can facilitate evaluation of the effects of new antiviral interventions within the maternal-fetal interface milieu.

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Section: Cells and Virusesmentioning

confidence: 99%

Modeling of Human Cytomegalovirus Maternal-Fetal Transmission in a Novel Decidual Organ Culture

Weisblum

Panet

Zakay‐Rones

et al. 2011

J Virol

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“…Normal and tumor tissues of human and mouse (BALB/c male mice [6 to 7 weeks old]) origin were washed five times in supplemented hormonal epithelial medium (SHEM) (the normal mouse colon lumen was first opened with scissors, and tissues were cut in a microtome [TC-2 tissue sectioner; Sorvall Corp.] into 500-m-thick slices). Tissues were incubated at 37°C in 5% CO 2 (5 slices/well in a 48-well plate) (14,22) in SHEM. The organ culture medium was SHEM (5), consisting of a 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium with 5% FCS, 10 mM HEPES, 0.5% dimethyl sulfoxide, 0.5 g/ml hydrocortisone, 1% minimal essential medium vitamin solution (Biological Industries, Beit Haemek, Israel), 5 g/ml insulin, 5 g/ml transferrin, 5 ng/ml selenium, 2 ng/ml epidermal growth factor, 100 U/ml penicillin, 100 g/ml streptomycin, 15 g/ml gentamicin, and 15 g/ml ciprofloxacin (Ciproxin).…”

Section: Cellsmentioning

confidence: 99%

“…Such a system may serve as a means to study viral tropism in the context of a three-dimensional tissue structure (8,14). Organ cultures derived from normal liver tissue and from ovarian and breast carcinomas have also been applied to the study of gene transfer by viral vectors (21,41,46,47).…”

mentioning

confidence: 99%

Herpes Simplex Virus Type 1 Preferentially Targets Human Colon Carcinoma: Role of Extracellular Matrix

Kolodkin-Gal

Zamir

Edden

et al. 2008

J Virol

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Viral therapy of cancer (viral oncolysis) is dependent on selective destruction of the tumor tissue compared with healthy tissues. Several factors, including receptor expression, extracellular components, and intracellular mechanisms, may influence viral oncolysis. In the present work, we studied the potential oncolytic activity of herpes simplex virus type 1 (HSV-1), using an organ culture system derived from colon carcinoma and healthy colon tissues of mouse and human origin. HSV-1 infected normal colons ex vivo at a very low efficiency, in contrast to high-efficiency infection of colon carcinoma tissue. In contrast, adenoviral and lentiviral vectors infected both tissues equally well. To investigate the mechanisms underlying the preferential affinity of HSV-1 for the carcinoma tissue, intracellular and extracellular factors were investigated. Two extracellular components, collagen and mucin molecules, were found to restrict HSV-1 infectivity in the healthy colon. The mucin layer of the healthy colon binds to HSV-1 and thereby blocks viral interaction with the epithelial cells of the tissue. In contrast, colon carcinomas express small amounts of collagen and mucin molecules and are thus permissive to HSV-1 infection. In agreement with the ex vivo system, HSV-1 injected into a mouse colon carcinoma in vivo significantly reduced the volume of the tumor. In conclusion, we describe a novel mechanism of viral selectivity for malignant tissues that is based on variance of the extracellular matrix between tumor and healthy tissues. These insights may facilitate new approaches to the application of HSV-1 as an oncolytic virus.

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“…The encapsulation strategy is even more compelling were ex vivo gene therapy of the implanted cells may be required [28]. This may have broader implications: for instance the skin MOs could be batch prepared from a particular source, genetically modified [17], tested for production of the target gene, cryopreserved and then encapsulated and implanted back into non-autologous hosts.…”

Section: Discussionmentioning

confidence: 99%

“…MOs are organfragments of microscopic thickness which retain the basic organ architecture and that ensure that no cell is more than 150 µm away from a source of nutrients and gases and thus remain viable for long periods even when cultured in serum-free medium. Lung and skin MOs have been characterized previously and found to remain viable and to transcribe tissue-specific genes for several weeks in culture in serum-free medium [17, 18]. …”

Section: Introductionmentioning

confidence: 99%

Skin-Derived Micro-Organs Induce Angiogenesis in Rabbits

Hasson

Gallula²,

Shimoni³

et al. 2006

J Vasc Res

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We have recently reported an alternative cell therapy approach to induce angiogenesis. The approach is based on small organ fragments – micro-organs (MOs) – whose geometry allows preservation of the natural epithelial/mesenchymal interactions and ensures appropriate diffusion of nutrients and gases to all cells. We have shown that lung-derived MOs, when implanted into hosts, transcribe a wide spectrum array of angiogenic factors and can induce an angiogenic response that can rescue experimentally induced ischemic regions in mice.From a clinical perspective, skin-derived MOs are particularly appealing as they could readily be obtained from a skin biopsy taken from the same target patient. In the present work we have investigated the angiogenesis-inducing capacity of rabbit and human skin-derived micro-organs in vitro and in vivo. Rabbit skin MOs were implanted into homologous adult rabbits and human skin MOs were encapsulated and implanted into xenogenic mice. Skin-derived MOs, as lung-derived MOs, were found to secrete a whole array of angiogenic factors and to induce a powerful angiogenic response when implanted back into animals. We believe the approach presented suggests a novel, efficacious and simple approach for therapeutic angiogenesis.

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Solid tissues can be manipulated ex vivo and used as vehicles for gene therapy

Cited by 30 publications

References 41 publications

Modeling of Human Cytomegalovirus Maternal-Fetal Transmission in a Novel Decidual Organ Culture

Modeling of Human Cytomegalovirus Maternal-Fetal Transmission in a Novel Decidual Organ Culture

Herpes Simplex Virus Type 1 Preferentially Targets Human Colon Carcinoma: Role of Extracellular Matrix

Skin-Derived Micro-Organs Induce Angiogenesis in Rabbits

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