Cell surface changes in a cell line derived from a human pleural effusion

The phenomenon of blebbing has been recorded in cells obtained from a pleural effusion of a patient with a small cell lung carcinoma. The blebbing manifested itself as a sudden ballooning of the cell membrane which then moved around the perimeter of the cell as a sausage-shaped swelling. This was followed by its rapid collapse. The rapidity of the process allowed real time video recordings, not time lapse, and still frames from the recording were photographed. Cells were also viewed using a scanning electron microscope and more detail of the blebs was revealed. Possible functions of cell blebbing are discussed.     

Establishment and characterization of HER2-positive cell line derived from pleural effusion of human breast scirrhous carcinoma

We have established a cell line derived from a pleural effusion of breast scirrhous carcinoma. This cell line presented immunohistochemically negative for estrogen receptor and slightly positive for progesterone receptor, and positive for c-erb B/HER2/neu. However the original tumor was found to be positive for estrogen receptor and progesterone receptor and slightly positive for c-erbB/HER2/neu expression. Enzyme and electrochemiluminescence immunoassays of tumor markers in the conditioned medium by the cell line revealed they secrete CA1 5-3, NCC-ST-439 and HER2 protein. We believe the cell line will contribute to the therapeutic study of malignant breast cancer.     

Establishment and characterization of a cell line (BTIC) including HER-2-positive cells derived from pleural effusion of recurrent breast invasive ductal carcinoma, scirrhous type

Human epidermal growth factor receptor-2 (HER-2) overexpression in breast cancer occurs in 20% to 30% of patients with breast cancer. Trastuzumab (Herceptin) targets HER-2 tyrosine kinase receptors expressed on tumor cells and mediates anti-proliferative effects against HER-2-positive tumor cells. Adjuvant chemotherapy with trastuzumab has improved the prognosis of patients with HER-2 positive high-grade breast cancer. However, patients often experience appearance and proliferation of recurrent tumor cells after trastuzumab treatment. In this study, we report the successful establishment and characterization of a cell line (BTIC) derived from a patient with recurrent breast cancer after adjuvant chemotherapy with trastuzumab. Characteristics of the BTIC cell line were investigated by phase contrast or electron microscopic observations, chromosome analysis, xenotransplantation, immunohistochemistry and radioimmunoassay for tumor markers. We confirmed that the BTIC cell line grown as multilayered culture in culture dishes, has a poorly developed endoplasmic reticulum in the cytoplasm and some desmosomes. The population doubling time was approximately 44 hr. A graft in nude mouse after xenotransplantation was diagnosed as scirrhous carcinoma. Immunohistochemistry on cultured BTIC cells revealed that the BTIC cells were negative for estrogen receptor and progesterone receptor, and 30% positive for HER-2. Radioimmunoassay indicated secretion of HER-2 protein, NCC-ST-439 and CA15-3.     

Establishment of a cell line derived from top-minnow (Gambusia patruelis)

A cell line, derived from top-minnow (Gambusia patruelis), has been transferred for 114 times in Leibovitz L-15 medium plus 10% FCS (GM-L-15) at a temperature of 31 degrees C and designated as TM. The results of a chromosome analysis of 100 cells at the metaphase stage, show a bimodal distribution of the chromosome number ranged from 21 to 68 with a modal number of 38. At a low seeding density, the plating efficiency of TM cells was 11-12.4%. TM cells are susceptible to EVE, EVA, EVEX, IPNV and LV-1 at 18 degrees C. The result of a sterility test showed that the TM cell line was free of bacterial, fungal and mycoplasmic contamination. The presence of numerous microvilli on the surface of TM cells in the micrographs of normal TM cell. The microvilli are replaced by blebs at the late stage of cytokinesis of the cells.     

Establishment and characterization of a primary and a metastatic melanoma cell line from Grey horses

The Grey horse phenotype, caused by a 4.6 kb duplication in Syntaxin 17, is strongly associated with high incidence of melanoma. In contrast to most human melanomas with an early onset of metastasis, the Grey horse melanomas have an extended period of benign growth, after which 50% or more eventually undergo progression and may metastasize. In efforts to define changes occurring during Grey horse melanoma progression, we established an in vitro model comprised of two cell lines, HoMel-L1 and HoMel-A1, representing a primary and a metastatic stage of the melanoma, respectively. The cell lines were examined for their growth and morphological characteristics, in vitro and in vivo oncogenic potential, chromosome numbers, and expression of melanocytic antigens and tumor suppressors. Both cell lines exhibited malignant characteristics; however, the metastatic HoMel-A1 showed a more aggressive phenotype characterized by higher proliferation rates, invasiveness, and a stronger tumorigenic potential both in vitro and in vivo. HoMel-A1 displayed a near-haploid karyotype, whereas HoMel-L1 was near-diploid. The cell lines expressed melanocytic lineage markers such as TYR, TRP1, MITF, PMEL, ASIP, MC1R, POMC, and KIT. The tumor suppressor p53 was strongly expressed in both cell lines, while the tumor suppressors p16 and PTEN were absent in HoMel-A1, potentially implicating significance of these pathways in the melanoma progression. This in vitro model system will not only aid in understanding of the Grey horse melanoma pathogenesis, but also in unraveling the steps during melanoma progression in general as well as being an invaluable tool for development of new therapeutic strategies.     

Establishment and characterization of a novel human endometrial mesenchymal stem cell line from a patient with adenomyosis

Adenomyosis, previously termed “endometriosis interna,” is a widespread disease affecting the female reproductive system and frequently resulting in infertility in women. The aim of this work was to examine the properties of endometrial mesenchymal stem cells (eMSCs) from a patient with adenomyosis. We established the cell line from a patient with adenomyosis and compared the properties of these cells with cells derived from a healthy donor. It was found that patient-derived eMSCs and eMSCs from healthy donors had a fibroblast-like morphology and did not differ in expression of surface markers and adipogenic potential. Karyotype analysis of G-banded metaphase chromosomes was performed in cells of both lines at the six or seventh passages. Cells from the healthy donor mostly had normal karyotype. Karyotype of eMSCs from the patient with adenomyosis usually had chromosomal abnormalities. The abnormalities concerned aneuploidy and nonrandom chromosomes breaks more often involving chromosomes 7 and 11. Although karyotype instability may be a sign of cell transformation, the patient-derived eMSCs stopped cycling after about 26 passages and entered into replicative senescence. This shows that the karyotypic abnormalities that we observed in adenomyosis-derived eMSCs are not relevant to the cell transformation and immortalization in vitro.     

Establishment and characterization of a bi-phenotypic leukemic cell line, NALM-19, from a patient with acute leukemia.

Based on the immunophenotypic and genotypic findings, this acute leukemia cell line, designated NALM-19, is unique in that a partial expression of both B-cell and myeloid cell features are present in this single clonal leukemic cell population. It is noteworthy that two "normal" EB virus-transformed B cell lines, B239 and B240, (paired with NALM-19) were established from the same leukemic blood.     

Establishment and characterization of a cell line (SS78) from a human renal cell carcinoma

Summary A new cell line, SS78, was established from a primary renal cell carcinoma of a Caucasian male. The tissue was dispersed with collagenase, and viable cells were separated by flotation on a Ficoll-Hypaque gradient. In culture, the SS78 cells retained a distinct epithelial morphology, and no fibroblastlike cells were seen. The cultured cells were aneuploid with a modal chromosome number of 80 and had several marker chromosomes. Inoculation of the cultured cells into athymic nude mice caused tumors at the sites of inoculation. This research was supported in part by Grants CA 15972 and CA 14930 from the National Cancer Institute through the National Bladder Cancer Project and by the Medical Research Service of the Veterans Administration.     

Establishment of a B-lymphoblastoid cell line infected with Epstein-Barr-related virus from a cynomolgus monkey (Macaca fascicularis)

A B-lymphoblastoid cell line (Ts-B) was established from lymph nodes of an apparently healthy cynomolgus monkey. The cells were demonstrated to contain Epstein-Barr virus-related antigens. Moreover, herpesvirus particles were found in the cells by electron microscopy. The cell-free culture medium transformed lymphocytes of cynomolgus rhesus monkeys, and of Japanese monkeys.     

Establishment and characterization of a malignant lymphoid cell line from a chronic lymphocytic leukemia patient

A long-term culture Epstein-Barr virus (EBV)-negative malignant lymphoid cell line (NAK) was established from a lymph node biopsy of a chronic lymphocytic leukemia patient. This cell line is of particular interest because it grows as an adherent cell line and depends on the presence of autologous conditioned medium for growth. After 6 months of growth in vitro, doubling time and cell cycle parameters were derived. Doubling time was 48 hours with over 45% cycling cells. Cell viability was over 90%. Expression of B-cell markers (CD19 and CD20) and surface immunoglobulin of the original tumor cell biopsy were roughly the same as in passage 14 (3 months in culture), including the expression of the original patient idiotype and IgM-lambda. Furthermore, binding of antiidiotypic antibodies was only slightly decreased at passage 14. Cytogenetic studies of chromosomal abnormalities in the primary tumor tissue and in later passages indicated similar abnormalities, with no translocations t(8;14), t(14;22), or t(2;8). However, frequent trisomies, deletions, and t(1;4) translocations were observed. Negative results for EBV nuclear antigen indicate that this cell line is an EBV-negative cell line.     

Establishment and characterization of a novel megakaryoblastic cell line, MOLM-1, from a patient with chronic myelogenous leukemia.

Based on the morphological resemblance to megakaryocytes and upon the expression f the platelet specific glycoprotein antigen, CD61, it is highly l y that MOLM-1 is a clonal leukemic cell population of megakaryoblastic origin .