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P Kaaijk

Publications and source records attributed to P Kaaijk.

9 recordsLinked to original sources

Differential expression of CD44 splice variants in the normal human central nervous system.

Cluster of differentiation 44 (CD44) is a broadly distributed group of glycoproteins that are involved in many functions related to cell-cell and cell-matrix interactions. In the present study, the expression of the standard form of CD44 (CD44s) and of CD44 variants (CD44v) was explored immunohistochemically on frozen sections of various areas of the human CNS. The results demonstrate that CD44s epitopes are expressed predominantly by white matter astrocytes, whereas different CD44 variant molecules are present in neurons, on axonal membranes, on endothelium or on choroid plexus epithelium. Interestingly, neurons and axons differentially expressed CD44 variant epitopes but consistently lack immunoreactivity for CD44s epitopes. Another interesting finding was that some CD44 variant epitopes expressed by neurons were localized in the cytoplasm instead of on the cell membrane. The broad distribution of variant CD44 molecules in the human CNS suggests that CD44 may play an important role in many biological processes in the CNS.

Aged↗

Hypofractionated radiation induces a decrease in cell proliferation but no histological damage to organotypic multicellular spheroids of human glioblastomas.

The aim of this study was to examine the effect of radiation on glioblastoma, using an organotypic multicellular spheroid (OMS) model. Most glioblastoma cell lines are, in contrast to glioblastomas in vivo, relatively radiosensitive. This limits the value of using cell lines for studying the radiation effect of glioblastomas. The advantage of OMS is maintenance of the characteristics of the original tumour, which is lost in conventional cell cultures. OMS prepared from four glioblastomas were treated with hypofractionated radiation with a radiobiologically equivalent dose to standard radiation treatment for glioblastoma patients. After treatment, the histology as well as the cell proliferation of the OMS was examined. After radiation, a significant decrease in cell proliferation was found, although no histological damage to the OMS was observed. The modest effects of radiation on the OMS are in agreement with the limited therapeutic value of radiotherapy for glioblastoma patients. Therefore, OMS seems to be a good alternative for cell lines to study the radiobiological effect on glioblastomas.

Cell Division↗

Daunorubicin and doxorubicin but not BCNU have deleterious effects on organotypic multicellular spheroids of gliomas.

In the present study organotypic multicellular spheroids (OMS) were used to study the effects of chemotherapeutic agents on malignant gliomas. Compared with the frequently used cell line models, OMS have several advantages with respect to the preservation of the cellular heterogeneity and the structure of the original tumour. OMS prepared from seven glioma specimens were treated with 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), daunorubicin or doxorubicin. After exposure to these drugs, the histology and cell proliferation of the OMS were analysed by immunohistochemistry and image analysis. Furthermore, the expression of P-glycoprotein (P-gp) and multidrug resistance-related protein (MRP), which both can contribute to resistance to daunorubicin and doxorubicin, were immunohistochemically investigated. We found that OMS from gliomas are sensitive for daunorubicin and doxorubicin but not for BCNU in terms of tissue destruction and decrease in cell proliferation. In addition, all gliomas were P-gp and MRP negative, which is in accordance with the sensitivity for daunorubicin and doxorubicin. Considering the potential use of several new alternative drug delivery methods, such as intratumoural implantation of drug-impregnated polymers or liposomal encapsulation of cytostatic drugs, daunorubicin and doxorubicin might be effective in the treatment of malignant gliomas.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Cryopreservation of organotypic multicellular spheroids from human gliomas.

Fresh human glioma tissue can be cultured on agarose to form organotypic multicellular spheroids (OMS). The major advantage of OMS is the preservation of the cellular heterogeneity and the tumour architecture, which is lost in conventional monolayer cultures. The present study was undertaken to assess the possibilities of storing frozen OMS from seven gliomas which were frozen to determine the viability after thawing. OMS were frozen slowly to -196 degrees C using a programmable freezing machine in culture medium containing 45% serum and 10% of the cryopreservative agent dimethyl sulphoxide (DMSO). After 2 weeks storage at -196 degrees C, quick thawing, and culturing for another week, it appeared that the frozen-thawed OMS were viable and retained their histological characteristics. In addition, it is demonstrated that the cellular constituents of the OMS resumed metabolic and proliferative activities. It is concluded that it is possible to establish frozen stocks of viable glioma OMS. This will enable extensive studies on OMS, such as investigation of the biological behaviour of gliomas by using OMS obtained from primary and corresponding recurrent gliomas. In addition, cryopreservation of OMS makes it possible to correlate the results of in vitro tests on OMS with the patients' responses to similar therapeutic approaches.

Brain Neoplasms↗

Expression of CD44 splice variants in human primary brain tumors.

Expression of CD44, particularly of certain splice variants, has been linked to tumor progression and metastatic potential in a number of different animal and human cancers. Although differential expression of CD44 standard epitopes (CD44s) in human brain tumors has been reported, the expression of CD44 variant exon encoded sequences (CD44v) in primary brain tumors in situ has not been studied in detail. In the present study, the expression of CD44s and CD44v epitopes was analyzed immunohistochemically on frozen sections of primary brain tumors. In addition, the expression of CD44 on cultured glioma cells was investigated by immunofluorescence flow cytometry. The results demonstrate the presence of CD44s epitopes and of CD44 splice variants containing CD44v4, v5 and v10 sequences in various types of brain tumors. A subgroup of highly malignant gliomas showed a strong (focal) expression of CD44v5. CD44v6 was absent in all brain tumors examined. CD44s appeared to be the dominant form of CD44 expressed in primary brain tumors, its expression was not correlated with tumor grade. We envisage that CD44 isoforms, in particular CD44s, may contribute to the invasive character of primary tumors by interacting with hyaluronate, one of the most abundant molecules in the extracellular matrix of the brain.

Alternative Splicing↗

Long-term culture of organotypic multicellular glioma spheroids: a good culture model for studying gliomas.

Gliomas, as well as other solid tumours, contain tumour stroma composed of connective tissue, macrophages, capillaries and other non-cellular constituents. Therefore, a homogeneous culture of tumour cells alone, as is often used as a culture model for gliomas, is not ideal to study all aspects of gliomas. In the present study we describe an alternative culture model, i.e. organotypic multicellular spheroids (OMS), that histologically closely resembles the tumour in vivo. Glioma explants, obtained at surgery from five patients, were cultured on agarose to form OMS, which were cultured for up to 16 weeks. At regular intervals, OMS were fixed and histological and immunocytochemical analyses were carried out. The histology as well as the immunocytochemical characteristics of the OMS proved to be almost unchanged after a culture period of 16 weeks. In contrast to monolayer cultures, glial fibrillary acidic protein (GFAP) expression in the OMS is preserved after 16 weeks of culture. However, in OMS from three out of five patients, small GFAP-negative cells appeared in the outer cell layers between 1 and 2 weeks of culture. Furthermore, after about 6 weeks of culture, the capillaries disappeared from the OMS. After prolonged culture, tumour cell heterogeneity, the cellular composition, and the histology of the OMS still closely resembled the tumour in vivo. It is suggested that OMS provide a good long-term culture model for the study of gliomas.

Adult↗

Cytolytic effects of autologous lymphokine-activated killer cells on organotypic multicellular spheroids of gliomas in vitro.

Knowledge about lymphokine-activated killer (LAK) cell infiltration and LAK cell cytotoxicity is essential to improve the effectiveness of LAK cell therapy against gliomas. In the present study, organotypic multicellular spheroids (OMS) of glioma tissue were used as a culture model to study the effects of LAK cells on gliomas. Compared to tumour cell lines and spheroids derived from tumour cell lines, OMS have several advantages with respect to preservation of tumour cell heterogeneity and the maintenance of the tumour architecture, e.g. capillaries and extracellular matrix. Four glioma specimens, obtained at surgery, were cultured directly on agarose to form OMS, which were then co-cultured with either autologous LAK cells or autologous non-activated peripheral blood lymphocytes (PBLs). After various time periods of co-cultivation, the OMS were fixed and examined both histologically and immunocytochemically. The present results showed that LAK cells infiltrated the OMS completely within 24 h of co-cultivation and severe cellular damage was observed, whereas PBLs infiltrated the OMS poorly and there was only marginal cellular damage. The present study indicates that OMS of gliomas provide an experimental model to investigate the infiltration and cytotoxicity of LAK cells on glioma tissue in vitro.

Adult↗

Overexpression of a M(r) 110,000 vesicular protein in non-P-glycoprotein-mediated multidrug resistance.

A M(r) 110,000 protein (p110) is overexpressed in P-glycoprotein-negative multidrug-resistant tumor cell lines of different histogenetic origins. These cell lines show an ATP-dependent drug accumulation defect, suggesting the presence of drug transporter molecules different from P-glycoprotein. Immunohistochemical staining with a p110-specific monoclonal antibody (LRP-56) showed that, like P-glycoprotein, the molecule has a high expression in normal epithelial cells and tissues chronically exposed to xenobiotics and potentially toxic agents, such as bronchial cells, cells lining the intestines, and kidney tubules. Staining of LRP-56 is primarily cytoplasmic, in a coarsely granular fashion, indicating that it reacts with a molecule closely associated with vesicular/lysosomal structures. Involvement of p110 in the energy-dependent drug transport process present in the cell lines is unknown.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

In vitro and in vivo models for the study of brain tumour invasion.

Since it is difficult to study the dynamic biological aspects of brain tumour invasion using histological sections of surgical biopsy and autopsy tissues, various laboratory systems have been devised. Animal models are less than ideal as chemically-induced brain tumours suffer from the fact that they have a low incidence and a long latency, while transplanted tumours grow predominantly by expansion, due to high proliferative activity, and not by diffuse local invasion as in human brain tumours. Various in vitro assays have, therefore, been established for both migration and invasion. These include the simple scratch technique in a confluent cell monolayer, the use of cloning rings and the "Transwell" modified Boyden chamber technique. More complex, three-dimensional culture model systems have also been developed, using chick heart, optic nerve or reaggregated fetal brain as "targets" for the invasion of neoplastic glia. Each method has yielded important information on the mechanisms which underlie brain tumour invasion. Moreover, individual microenvironmental factors may be modulated in these laboratory systems to determine their influence on the migration/invasion process.

Animals↗