Establishment and characterization of a human liposarcoma cell line (HTLS) from the retroperitoneal liposarcoma

A cell line designated HTLS was established from the retroperitoneal liposarcoma. The HTLS line showed stable proliferation without interruption for 2 years and subcultivated over 35 times. The cells were elongated fibrous and spindle in shape, and neoplastic and pleomorphic features. The multinucleated giant cells with fine cytoplasm were seen. The cells proliferated slowly and the population doubling time was about 90 hours. The chromosome number showed a wide distribution of aneuploidy, the mode was hyperdiploid range (51-52), and many marker chromosomes were observed. The cells were transplantable into the submucosa of immunesuppressed hamster's cheek pouch and produced liposarcoma, while were not transplantable into subcutis of nude mice     

Karyology of primary human fetal cell cultures

Summary Cell cultures were established from the biopsies of lung, skin and kidney from each of nine human fetuses, and chromosome analyses were performed on material through the fifth subculture. Kidney cell cultures generally showed a higher level of polyploidy than lung or skin. The frequencies of hyperdiploid cells and those with structural abnormalities were consistent with the low levels found in cultures of human lymphocytes. The data provide a normal cytogenetic baseline for human fetal material which may be useful in a variety of studies. Supported by Food and Drug Administration contracts FDA 74-51 and 74-52. Authors are listed alphabetically.     

The human intestinal epithelial cell line Caco-2; pharmacological and pharmacokinetic applications

The gastrointestinal tract remains the most popular and acceptable route of administration for drugs. It offers the great advantage of convenience and many compounds are well absorbed and thereby provide acceptable plasma concentration-time profiles. Currently there is considerable interest from the pharmaceutical industry in development of cell culture systems that would mimic the intestinal mucosa in order to evaluate strategies for investigating and/or enhancing drug absorption. The intestinal epithelial cells of primary interest, from the standpoint of drug absorption and metabolism, are the villus cells, which are fully differentiated cells. Anin vitro cell culture system consisting of a monolayer of viable, polarized and fully differentiated villus cells, similar to that found in the small intestine, would be a valuable tool in the study of drug and nutrient transport and metabolism.The Caco-2 cell line, which exhibits a well-differentiated brush border on the apical surface and tight junctions, and expresses typical small-intestinal microvillus hydrolases and nutrient transporters, has proven to be the most popularin vitro model (a) to rapidly assess the cellular permeability of potential drug candidates, (b) to elucidate pathways of drug transport (e.g., passive versus carrier mediated), (c) to assess formulation strategies designed to enhance membrane permeability, (d) to determine the optimal physicochemical characteristics for passive diffusion of drugs, and (e) to assess potential toxic effects of drug candidates or formulation components on this biological barrier. Since differentiated Caco-2 cells express various cytochrome P450 isoforms and phase II enzymes such as UDP-glucuronosyltransferases, sulfotransferases and glutathione-S-transferases, this model could also allow the study of presystemic drug metabolism.     

Growth inhibitory effect of quercetin on SW 872 human liposarcoma cells

Adipocytic tumors represent the largest single group of soft tissue tumors. In the present study, we investigated the antiproliferative potential of quercetin in SW 872 human liposarcoma cells. Cell viability was significantly influenced by quercetin treatment in a time- and dose-dependent manner. Flow cytometric analyses of SW 872 human liposarcoma cells exposed to quercetin showed that the increase of apoptotic cells was time- and dose-dependent. The percentages of normal cells were decreased and apoptotic cells (including early apoptotic and late apoptotic) were increased with increasing concentrations of quercetin. Quercetin-induced apoptosis in SW 872 human liposarcoma cells was associated with the loss of mitochondrial membrane potential (DeltaPsi(m)). The apoptosis in SW 872 human liposarcoma cells induced by quercetin was mediated through the activation of caspase-3, Bax, and Bak and then cleavage of PARP and downregulation of Bcl-2. These results demonstrate that quercetin may prevent atypical lipomatous tumors/well-differentiated liposarcomas from mature adipocytic proliferation, which may contribute to its antiproliferative function.     

Chromosome studies on a human liposarcoma cell line.

Numerical and structural chromosome analysis of a human retroperitoneal liposarcoma cell line maintained under standard cell culture conditions revealed a very stable hypodiploid mode. If the cells were not trypsinized for several generations, a near-triploid stemline, which was generally a duplication of the hypodiploid mode, emerged. Some chromosomes appeared to be relatively stable pairs (1, 2, 7, 9, and 12), but most had "lost" one homolog or both (4 and 21) or were rearranged into "new" marker chromosomes. Quantitation of the genetic material showed a loss of 12.0 +/- 3.7% per spread. Only one characteristically long marker chromosome, which is present in every cell, could be identified with certainty as a translocation between chromosomes 4 and 11. Several of the marker chromosomes showed interstitial negatively staining regions with the trypsin-Giemsa method.     

A continuous cell line of a nonhematophagous mosquito,Toxorhynchites amboinensis

Summary A continuous cell line of a nonhematophagous mosquito was obtained by using partially incapacitated but living larvae as the source The cells were predominantly epithelioid and diploid (2N=6). Syncytia were occasionally observed in the normal cell cultures.     

Giant dedifferentiated liposarcoma of the right hemifacial area involving the oral cavity

Although liposarcoma is a reasonably common soft tissue sarcoma in adults, its occurrence within the head and neck region is very rare. The following report presents the case of a giant dedifferentiated liposarcoma initially located in the temporal region and then extending to the entire right maxillofacial region. Clinical as well as histopathological features and therapeutic approaches of dedifferentiated liposarcoma are discussed, and a literature review is presented.     

Characterization of newly established human osteosarcoma cell line,LM-1

Summary The characteristics of Cell Line LM-1, established from a human osteosarcoma, have been studied extensively. The cell produced both bone-specific and placental-like alkaline phosphatases when treated with hydrocortisone 21-phosphate; they had specific membrane antigens that reacted with sera from osteosarcoma patients. Injection of LM-1 cells into newborn hamsters treated with antilymphocyte serum produced nodular tumors. The characteristics of LM-1 suggest that this tumor cell line has unique features that may be useful in a variety of studies of human and animal osteosarcoma. This research is supported in part by USPHS Grants HD-09938 and CA-25746.     

A human gallbladder adenocarcinoma cell line

Summary A continuous cell line, COLO 346, was established from a liver metastasis in a patient with adenocarcinoma of the gallbladder. COLO 346 grew as an adherent monolayer of pleomorphic epithelioid cells. COLO 346 cells produced esterone, but no estradiol, progesterone, or cortisol. No adrenocorticotropic hormone, β-subunit of human chorionic gonadotropin, carcinoembryonic antigen, or α-fetoprotein production by the cells was detected. Cell doubling time was 36 h. Seven allelic isozymes were assayed. COLO 346 had a chromosome mode of 74 at 21 months postestablishment with 6 marker chromosomes present in 100% of the cells analyzed. COLO 346 has been in continuous culture for over 2 yr and is available to other investigators for their studies. This paper was presented in part at the 31 st Annual Meeting of the Tissue Culture Association, June 1–5, 1980. The work was supported by Grants CA15018 and CA29514 from the National Cancer Institute, and by the Mary B. and L. H. Marshall Fund.     

Short-term cell culture technique for obtaining chromosomes in marine and freshwater fish

In this paper we describe a simple and reliable short-term fish cell culture technique, for obtaining chromosome spreads with minimum stress involving the specimens sampled. Fibroblasts were derived from primary cultures from the caudal fin of some economically important freshwater species: northern pike, Esox lucius L. , goldfish, Carassius auratus L., and marine species: gilthead sea bream, Sparus aurata L. and cardinal fish, Apogon imberbis L.     

Clinico-molecular study of dedifferentiation in well-differentiated liposarcoma

Well-differentiated liposarcoma (WD) acquires fully malignant potential when the histological progression named dedifferentiation occurs. This progression is supposed to occur in a time-dependent manner but this is still a debated issue. Clinically, the prediction of dedifferentiation for WD is very important from the therapeutic point of view. To identify genes that are predictive of dedifferentiation and to understand the mechanism of dedifferentiation, we investigated clinical information of 50 cases and studied the gene expression profiles of 36 lipomatous tumors using cDNA microarray. The clinical study showed that the dedifferentiation did not always seem to occur in a time-dependent manner. Interestingly, from the gene expression study, unsupervised hierarchical clustering analysis of well-differentiated lesions obtained from dedifferentiated liposarcoma (DD) cases that were indistinguishable from WD pathologically showed a clearly distinct gene expression pattern from WD. Using the pattern-matching program, 1687 genes including 487 known genes were identified, which discriminated WD cases from well-differentiated lipomatous lesions obtained from DD cases. These results suggest that the dedifferentiation may arise from different types of WD that could be distinguished from gene expression profiling but could hardly be classified by the pathological studies.     

Characteristics of seven clones of the WiDr human colon adenocarcinoma cell line

Summary Seven clonal populations were derived from the WiDr human colon adenocarcinoma cell line and were characterized with respect to chromosome number, DNA content, and tumorigenicity in nude mide. There was no correlation between tumor volume and either DNA content or chromosome number; but there were wide differences among the clones regarding the size of tumors they were able to produce in nude mice.     

A virus-producing cell line developed by transformation of human parapharyngeal cells with SV40

Summary Normal cultures of epithelial appearance were initiated by trypsinization of a surgically resected, histologically normal branchial cyst. Cellular morphology was consistent with derivation from the stratified squamous epithelium of the cyst or from vascular endothelium, although electron micrographs of the cultured cells failed to show any junctional complexes. Infection with SV40 produced transformants which were also epithelioid in appearance. These grew vigorously for 22 to 50 population doublings (about 23 to 32 subcultures, depending upon regimen) and then became quiescent. During this evolution, virus was detectable at all stages by both direct isolation (cell extracts) and cocultivation with permissive cells. In two sublines in which selection for rapidly growing cell types occurred, virus was detected only by cocultivation. The work confirms that of others in the finding that normal human epithelial cells are susceptible to transformation by oncogenic viruses, but are apparently less responsive than are fibroblasts to such transforming agents. It also suggests that subcultivation techniques that maintain the populations of transformed cells at low density tend to select against cell strains that are continuous producers of infectious virus. This study was supported by Grant CA 13494 from the National Cancer Institute.     

Characterization of a cell line derived from zebrafish (brachydanio rerio) embryos

Summary During the last decade, zebrafish (Brachydanio rerio) have emerged as a novel and attractive system to study embryogenesis and organogenesis in vertebrates. The main reason is that both extensive genetic studies and detailed embryologic analysis are possible using this small tropical fresh water teleost. However, in vitro analysis using cell culture or molecular genetics are still far less advanced than in other vertebrate systems. Here we report the generation and characterization of a fibroblast like cell line, ZF4, derived from 1-day-old zebrafish embryos. The hyperploid cell line has been stable in multiple passages for more than 2 yr now and is the first zebrafish cell line that can be maintained in conventional medium containing mammalian serum. Using a series of plasmids for expression of a marker gene, we evaluate in ZF4 cells the relative strength of expression from several different viral, fish, and mammalian promoters. Stable integration can be obtained by using G418 selection. We hope that our cell line will be a useful tool for the analysis of gene regulation in zebrafish.     

EPN: a novel epithelial cell line derived from human prostate tissue

This work reports the isolation and characterization of a line of human, nontransformed and differentiated prostate epithelial cells (EPN) in continuous culture. Primary cultures of epithelial prostate cells were set up using normal tissue isolated from a prostate sample collected after radical prostatectomy for cancer. After 70 passages, EPN cells did not undergo "Hayflike crisis" and were free of fibroblast contamination and were thus subcloned and characterized. EPN cells in culture, as prostate epithelial cells in vivo, express high-molecular weight cytokeratin and Pyk2, whereas they do not express desmin. EPN cells are nontransformed because they do not form colonies in semisolid medium and do not form tumors once injected into nude mice. EPN cells express the functional androgen receptor, which can mediate the mitogenic activity of testosterone. Finally, clonal production of the prostate-specific antigen could be detected in EPN cells. The availability of a line of epithelial nontransformed prostate cell in culture will be useful in investigating the complex process regulating normal prostate physiology as well as the development and progression of prostate tumors.     

Characterization of a reptilian epithelioid skin cell line derived from the green sea turtle,Chelonia mydas

Summary A continuous line of epithelioid cells was established from explant skin tissues of the green sea turtle,Chelonia mydas. These cells, designated GTS, have been subcultured more than 60 times in commercially available mammalian cell culture medium supplemented with 5% bovine calf serum. Of those temperatures tested, optimal growth was achieved at 30°C although replication occurred between 16 and 37°C. These cells may be held as monolayers at 8°C or stored frozen in growth medium containing 10% dimethylsulfoxide at −70 or −196°C. The modal number of 55 chromosomes per cell is in agreement with the heterogametic female diploid number of this species. The GTS line represents the first established culture of normal epithelioid skin cells to be reported for a poikilothermic species.     

A transitional cell carcinoma cell line

Summary A transitional cell carcinoma cell line, COLO 232, was derived from a primary urinary bladder tumor in a Caucasian male. In culture, COLO 232 retained distinct uroepithelial phenotypic traits and produced both carcinoembryonic antigen and adrenocorticotropic hormone. COLO 232 had a chromosome mode of 58 and retained the X and Y chromosomes. Ten marker chromosomes were identified. COLO 232 will be of value for biochemical and immunological studies. Presented in part at the 28th Annual Meeting of the Tissue Culture Association, June 7, 1977. This work was supported by Grant No. CA 15018 awarded by the National Cancer Institute, DHEW, and the Mary B. and L. H. Marshall Fund.     

A new parental cell line for human x human hybridoma production

The potential of a new HAT-sensitive human lymphoblastoid cell line TK6 TG^r.P1. as a fusion partner was assessed, by comparison with the established human parental cell line UC729.6. Both of these cell lines were fused with the peripheral blood mononuclear cells of a patient with B-chronic lymphocytic leukaemia. The hybridomas generated in these fusion experiments were analysed by the fluorescence activated cell sorter and karyotyping. An anti-idiotype ELISA assay detected the presence of the patient's characteristic idiotype bearing immunoglobulin in the supernatant of a number of the hybridoma cell lines generated in both fusions.     

Characterizations of the murine mesenchymal cell line expressing GFP

We established and characterized a murine mesenchymal stem cell line from the bone marrow of a transgenic C57BL mouse that ubiquitously expressed green fluorescent protein (GFP). Immunostaining revealed the presence of several markers common for mesenchymal stem cells (MSCs). The cells expressed specific fibroblast proteins, such as smooth muscle actin, which is localized in stress fibrils, and vimentin, a major protein of intermediate filaments in connective tissue cells. These proteins are responsible for the ability to differentiate into adipocytes or osteoblasts under appropriate conditions. The MSC karyotype was unstable. At the 6th passage cells, were aneuploid and genetically heterogeneous. The number of chromosomes ranged from near 2n to 8n. 80% of cells had chromosome numbers between 50 and 85 without a well-defined modal class. Differential G-staining of metaphase spreads showed variability in the copy numbers of individual chromosomes and presence of random chromosome rearrangements, such as ectopic associations of nonhomologous chromosomes. All cells analyzed contained a single dicentric marker chromosome. Some cells also had mini-chromosomes regarded as indicators of gene amplification. We suppose that the karyotypic instability of MSCs that express GFP is provoked by the insertion of foreign GFP transgenes into the murine genome. These cells could be useful for the study of genomic alterations during the spontaneous oncogenic transformation of stem cells.