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At least 145 records · Page 8Linked to original sources

Collagen matrix promotes reorganization of pancreatic endocrine cell monolayers into islet-like organoids.

To evaluate the capacity of pancreatic endocrine cells to reassociate in vitro according to the characteristic topographical pattern observed in the islets of Langerhans in situ, we cultured cells dissociated from neonatal rat pancreas within a three-dimensional collagen matrix. Cell monolayers grown on the surface of collagen gels were covered with a second layer of collagen. This induced the monolayers of endocrine cells to reorganize into smooth-contoured, three-dimensional aggregates, in which non-B cells (identified by electron microscopy and immunofluorescence) had a preferential distribution at the periphery, whereas B cells were concentrated in a central position. These results show that cultured pancreatic endocrine cells have the capacity to reassociate into islet-like organoids in vitro, and that collagen matrices may have a permissive effect on the expression of this potential.

Animals↗

Small intestinal organoid-derived SP cells contribute to repair of irradiation-induced skin injury.

Side population (SP) cells, characterized by their ability to efflux the fluorescent dye Hoechst 33342, were isolated from the small intestine of mice. In the abdominal irradiation model, small intestinal organoid-derived SP (sioSP) cells from ROSA 26 mice were submucosally injected into the small intestinal of the irradiated C57BL/6 mice. In contrast to the control mice, mice receiving sioSP cell transplantation demonstrated far less skin injury. Most importantly, hairs in the irradiated body part of the transplanted mice almost remained black, whereas the counterpart in the control mice almost turned white. Histochemistry studies showed the donor cells gave rise to skin cells in the irradiated skin. Thus, our study demonstrated for the first time that stem cells from the small intestine can differentiate into skin cells under local cues and thus supports the theory of stem cell plasticity.

Animals↗

Mucinous bronchioloalveolar carcinoma with organoid differentiation simulating the pyloric mucosa of the stomach: clinicopathologic, histochemical, and immunohistochemical analysis.

Seven cases of mucus-producing bronchioloalveolar carcinoma, which showed organoid differentiation simulating the gastric pyloric mucosa, were found among 176 cases of lung cancer. This type of adenocarcinoma, which corresponds to bronchioloalveolar carcinoma with mucus-secreting cells in the World Health Organization classification, characteristically formed papillary structures composed of two types of mucus cells: tall columnar cells in the upper portion of the papillary structure and more cuboidal cells in the lower portion. The former contained gastric surface mucous cell-type mucins that stained with galactose oxidase-cold thionine Schiff, whereas the latter possessed gastric gland mucous cell-type mucins specifically stained by paradoxical concanavalin A and were also positive for lysozyme and pepsinogen II by immunostaining. Chromogranin A-reactive tumor cells were also scattered among these tumor cells. This pattern of mucus-secreting cells, therefore, simulated the normal pyloric mucosa of the stomach.

Adenocarcinoma, Bronchiolo-Alveolar↗

Congenital apocrine hamartoma: an unusual clinical variant of organoid nevus with apocrine differentiation.

Congenital apocrine hamartomas are rare tumors of the skin composed of mature apocrine glands in the papillary and reticular dermis. Several cases have been reported in the literature but few were of uniform clinical appearance. The term apocrine nevus has been used interchangeably with apocrine hamartoma in the literature, however, based on their distinctive morphologies we believe they are different entities. We describe a 7-year-old girl with a congenital apocrine hamartoma of the left cheek. A stained biopsy specimen from the lesion revealed large mature apocrine glands with decapitation secretion in the dermis. The presence of periodic acid-Schiff-positive, diastase-resistant granular material in the apical cytoplasm of some secretory cells helped differentiate these as apocrine glands. We think this lesion may represent a form of organoid nevus with pure apocrine differentiation.

Child↗

Remodeling of resistance arteries in organoid culture is modulated by pressure and pressure pulsation and depends on vasomotion.

The hypothesis was tested that pressure and pressure pulsation modulate vascular remodeling. Arterioles ( approximately 200 microm lumen diameter) were dissected from rat cremaster muscle and studied in organoid culture. In the first series, arterioles were kept at a stable pressure level of either 50 or 100 mmHg for 3 days. Both groups showed a progressive increase in myogenic tone during the experiment. Arterioles kept at 50 mmHg showed larger endothelium-dependent dilation, compared with vessels kept at 100 mmHg on day 3. Remodeling, as indicated by the reduction in maximally dilated diameter at 100 mmHg, was larger in arterioles kept at 50 mmHg compared with 100 mmHg: 34 +/- 4.5 versus 10 +/- 4.8 microm (P < 0.05). In the second series, arterioles were subjected to a stable pressure of 60 mmHg or oscillating pressure of 60 +/- 10 mmHg (1.5 Hz) for 4 days. Pressure pulsation induced partial dilation and was associated with less remodeling: 34 +/- 4.0 versus 19 +/- 4.5 microm (P < 0.01) for stable pressure versus oscillating pressure. Vasomotion was frequently observed in all groups, and inward remodeling was larger in vessels with vasomotion: 30 +/- 2.5 microm compared with vessels that did not exhibit vasomotion: 8.0 +/- 5.0 microm (P < 0.01). In conclusion, these results indicate that remodeling is not enhanced by high pressure. Pressure pulsation causes partial dilation and reduces inward remodeling. The appearance of vasomotion is associated with enhanced inward remodeling.

Animals↗

Organoid culture of cannulated rat resistance arteries: effect of serum factors on vasoactivity and remodeling.

We developed an organoid culture technique to study the mechanisms involved in arterial remodeling. Resistance arteries were isolated from rat cremaster muscle and mounted in a pressure myograph at 75 mmHg. Vessels were studied during a 4-day culture period in DMEM with either 2% albumin, 10% heat-inactivated FCS (HI-FCS) or 10% dialyzed HI-FCS (12 kDa cut off) added to the perfusate. The albumin group showed a gradual loss of endothelial function and integrity, whereas smooth muscle agonist and myogenic responses were retained. No remodeling was observed. Vessels cultured in the presence of serum showed a progressive constriction. Smooth muscle responses and substance P-induced endothelium-dependent dilation were maintained. An inward remodeling of 17 +/- 4% in the HI-FCS group and 26 +/- 3% in the dialyzed HI-FCS group was found, while media cross-sectional areas were unchanged. These data show that pressurized resistance arteries can be maintained in culture for several days and undergo eutrophic remodeling in vitro in the presence of high molecular weight serum factors.

Acetylcholine↗

Connexins act as tumor suppressors in three-dimensional mammary cell organoids by regulating differentiation and angiogenesis.

Connexins are tumor suppressors, and human breast connexin 26 (Cx26) and connexin 43 (Cx43) gap junctions are often down-regulated in breast cancer. We previously showed that Cx26 and Cx43 overexpressed in MDA-MB-231 breast cancer cells inhibited tumor growth in vivo but not in two-dimensional cultures. In the current study, we show that overexpression of Cx26 or Cx43 has tumor-suppressive properties in a three-dimensional environment such that they reduced anchorage-independent cell growth and induced partial redifferentiation of three-dimensional organoids of MDA-MB-231 cells. Importantly, the majority of exogenous connexins did not localize to the cell-cell interface or rescue gap junctional intercellular communication (GJIC) as assessed by dye transfer, providing evidence of a GJIC-independent mechanism of mammary tumor suppression. To further elucidate the mechanisms involved in connexin-induced three-dimensional redifferentiation of tumor cells, we examined whether connexin expression has a role in epithelial to mesenchymal transition (EMT). Cx26 and Cx43 reduced cell migration, increased cytokeratin 18 expression, and decreased vimentin levels, indicating a shift from a mesenchymal towards an epithelial phenotype. In addition, we examined the role of connexins in angiogenesis by probing an angiogenesis antibody array with conditioned media from three-dimensional MDA-MB-231 cultures. This revealed that connexin overexpression regulated various angiogenesis-linked proteins. Furthermore, secreted factors from connexin overexpressing cells inhibited endothelial cell tubulogenesis and migration, and xenografts of Cx43 overexpressing MDA-MB-231 cells showed reduced tumor angiogenesis. In summary, Cx26 and Cx43 inhibit the malignant properties of MDA-MB-231 cells via GJIC-independent mechanisms, including regulation of EMT and angiogenesis.

Animals↗

A population of c-Kit(low)(CD45/TER119)- hepatic cell progenitors of 11-day postcoitus mouse embryo liver reconstitutes cell-depleted liver organoids.

Embryo liver morphogenesis takes place after gastrulation and starts with a ventral foregut evagination that reacts to factor signaling from both cardiac mesoderm and septum transversum mesenchyme. Current knowledge of the progenitor stem cell populations involved in this early embryo liver development is scarce. We describe here a population of 11-day postcoitus c-Kit(low)(CD45/TER119)- liver progenitors that selectively expressed hepatospecific genes and proteins in vivo, was self-maintained in vitro by long-term proliferation, and simultaneously differentiated into functional hepatocytes and bile duct cells. Purified c-Kit(low)(CD45/TER119)- liver cells cocultured with cell-depleted fetal liver fragments engrafted and repopulated the hepatic cell compartments of the latter organoids, suggesting that they may include the embryonic stem cells responsible for liver development.

Animals↗

Histologic types of thymoma associated with pure red cell aplasia: a study of five cases including a composite tumor of organoid thymoma associated with an unusual lipofibroadenoma.

The histologic type of thymomas associated with pure red cell aplasia (PRCA) has generally been regarded to be predominantly the spindle cell type. The 5 thymomas associated with PRCA we studied showed various histologic types and none of them was a spindle cell thymoma. Some cases reported as spindle cell thymoma associated with PRCA in the literature might not be spindle cell type. The discrepancy could be due to the lack of consensus in the histologic classification of thymomas in the past. Our study showed that thymomas associated with PRCA lacked a strong correlation with a particular histologic type and the pathogenesis of PRCA associated with thymoma did not seem to be related to the histologic type. One of our cases was an unusual composite tumor of an organoid thymoma (WHO type B1) and a hitherto undescribed "lipofibroadenoma." The clinical outcome was influenced by the invasive character and the histologic type of the thymoma.

Adult↗

Secretion of lamellar bodies in type II pneumocytes in organoid culture: effects of colchicine and cytochalasin B.

Lamellar bodies are the morphological correlate to the alveolar surface-active agent. Diminished synthesis of this surfactant in newborns results in the respiratory distress syndrome. Secretion of lamellar bodies and its dependence on the cytoskeleton are not yet well understood and are still controversially discussed in the literature. We therefore used an organoid culture system of fetal mouse lung cells to investigate electron microscopically secretion of lamellar bodies and influence of both colchicine and cytochalasin B on the secretion process. Secretion of lamellar bodies is to be considered as an eccrine secretion, because no other cytoplasmic components were extruded. It includes a very fast component in opening of the secretory vacuole. Synthesis, but not secretion, was inhibited by colchicine; secretion, but not synthesis, was inhibited by cytochalasin B. Furthermore, formation of the histotypic structures in vitro and deposition of the basal lamina were disturbed by cytochalasin B, but not altered by colchicine. After short-term treatment, these effects turned out to be reversible. The results indicate that synthesis of lamellar bodies depends on an intact microtubular system, whereas secretion requires actin filaments in a functional state.

Animals↗

Growth and morphogenesis of epithelial cell organoids from peripheral and medial mammary parenchyma of prepubertal heifers.

Collagen gel cell cultures were used to test the hypothesis that cells from peripheral parenchymal zones of the developing bovine mammary gland have greater proliferative and morphogenic potential than cells from the medial parenchymal mass. Tritiated thymidine was incorporated to assess cell proliferation. Dose responses for insulin-like growth factor one (IGF-I, 0 to 50 ng/ml) and transforming growth factor beta one (0 to 5 ng/ml) were determined for each cell population. Cells from the peripheral regions were two to three times more sensitive to the mitogenic action of IGF-I or serum. Transforming growth factor beta one concentrations of 12.5 to 500 pg/ml stimulated cell proliferation, but concentrations above 1 ng/ml were inhibitory. Peripheral cells showed a distinct biphasic response to addition of transforming growth factor beta one. Morphology of organoids was evaluated daily during culture and by histologic examination at the end of culture. Peripheral zone cells formed multi-layered cell structures that resembled native mammary parenchyma with serum or mammary gland extract. Our results indicate that epithelial cells of the peripheral and medial parenchymal zones in the ruminant mammary gland can differ in their proliferative and morphogenetic response to growth factor treatment in vitro. These differences likely reflect local tissue regulation necessary for sequential ductular and lobulo-alveolar development in vivo.

Animals↗

Atypical oculo-orbital complex choristoma in organoid nevus syndrome.

We report organoid nevus with left oculo-orbital choristoma fixing the globe in hypotropia, precanthal skin tag, and upper eyelid coloboma and right limbal dermoid. Computed tomography revealed a dilated lateral ventricle and cerebral atrophy. No neurologic deficit existed. The coloboma was repaired, choristoma debulked, and precanthal tag excised.

Calcinosis↗

[Organoid nevus (nevus sebaceus) dominated by the apocrine glands].

We report a case of congenital organoid nevus with an excessive number of apocrine glands. A one-year-old boy presented with a slowly enlarging keratotic element in the left temporal region, and biopsy showed hyperkeratosis, acanthosis, follicular plugging and multiple transverse sections of mature apocrine glands filling the reticular dermis. The lesion was benign and treatment was therefore conservative. In rare cases, various types of adnexal neoplasms may develop, but malignant tumours occur very seldom.

Apocrine Glands↗

An unusual organoid nevus.

The sites of predilection, stages of development, and suggested therapy for organoid nevi are discussed herein, and the case of an unusual nevus is presented.

Adolescent↗

Human colon adenocarcinoma cells. III. In vitro organoid expression and carcinoembryonic antigen kinetics in hollow fiber culture.

Cell line LS174T was established in our laboratory from a primary human colon adenocarcinoma and was serially cultured for 4 years. Following 1 month of culture in perfused hollow fiber matrices, organoid growth reminiscent of the patient's original tumor was observed. The cellular organization was predominantly glandular with mucin located within gland lumina and cells. Glands had irregular brush borders, well-developed junctional complexes, and intracytoplasmic lumina lined with microvilli, features found in adenocarcinoma tissue. Alternate-day determinations of carcinoembryonic antigen (CEA) revealed biphasic kinetics in the extracapillary fluids of the hollow fiber system but not in identically treated monolayer cultures. The initial rate of CEA release was similar to that of monolayer cultures. A significantly accelerated secondary CEA release phase was observed in 10,000- and 50,000-molecular weight (MW)-exclusion fiber cultures (P less than or equal to 0.001 and P = 0.05, respectively). Totals of 43.8 micrograms and 112.6 micrograms CEA were released into the extracapillary fluids of 10,000- and 50,000-MW-exclusion fiber cultures, respectively, which were onefold and 2.3-fold increases, respectively, over monolayer supernatant yields. Most CEA was released during the final days of the secondary phase. In monolayer cultures, maximum CEA release occurred during the stationary phase of growth.

Adenocarcinoma↗

Synovial and peritoneal macrophages in organoid culture.

Cultivation of macrophages and their progenitors has been very useful for elucidation of function, behaviour and morphology of these cells. The purpose of this contribution is to describe a new in vitro system (organoid, high density or micromass culture) which proved to be convenient for cultivation of macrophages derived from human synovial fluid and tissue and mouse peritoneal fluid. Using this method, highly differentiated and functionally active macrophages of marked purity and long maintenance (up to 2 weeks) could be obtained even after previous cultivation and subcultivation in monolayer culture. The macrophages were identified by electron microscopy and immunomorphology using HLA-DR-DP, CD-68 (markers for human macrophages), anti-human-polymorphonuclear leukocyte-gelatinase and F4/80 (a mouse macrophage surface marker). The significance of this method as a research tool in the study of cartilage degradation by macrophages in co-cultures is stressed.

Animals↗

Matrix changes during long-term cultivation of cartilage (organoid or high-density cultures).

In high density (organoid or micromass) cultures of prechondrogenic mesenchymal cells from limb buds of 12-day-old mouse embryos typical cartilaginous tissue develops after 3 days. Immunomorphological investigations have shown that it contains the typical components of the cartilaginous matrix, such as collagen type II and cartilage-specific proteoglycans. After a 2-week cultivation period hypertrophic cartilage cells develop to an increasing extent. Many of these cells as well as normal chondroblasts detach from the matrix from the 2nd week in vitro onwards to assume a fibroblast-like appearance. At the same time thick (25-65 nm) collagenous fibrils occur at the surface of these cells. These thick fibrils contain collagen type I, as shown by immunomorphology. Hence, in these older cartilage cultures chondroblasts change their synthesis programme or direction of differentiation. Consequently, a model for the study of "dedifferentiation" of cartilage and possibly also transformation of cartilage cells to osteoblasts has become available.

Animals↗

[Mixed "organoid" tumor or the testis: general considerations. Report of a case].

Presentation of one case of mixed testicular tumour in a 31-year-old male patient. Since this is the association in the same organ of an "organoid" teratoma plus a seminoma, the histological diversity, frequency, as well as the analysis of the controversies and criteria for its therapeutical approach are all discussed. Also, several histological theories on the genesis of mixed testicular tumours are explained. Prognosis and evolution after treatment of the present case is included.

Adult↗