Chieko Ishiwata

Establishment of human endometrial adenocarcinoma cell line containing estradiol-17β and progesterone receptors

The cell line designated HHUA was established from a well-differentiated endometrial adenocarcinoma (grade 1) of a 54-year-old female on December 10, 1980. The cell line grew well without interruption for 22 months and was subcultivated more than 56 times. The cells were spindle or polygonal in shape, formed multilayers, and did not show contact inhibition. The chromosome number varied widely and showed aneuploidy. The modal number was 46 and no marker chromosome was identified. The cells were transplanted into the subcutis of BALB/C nude mice and produced tumors which were interpreted as adenocarcinoma. The cells contained estradiol-17 beta and progesterone receptors. Administration of estradiol-17 beta successfully depleted cytoplasmic estrogen receptors and brought about increased progesterone receptor levels.

Establishment and characterization of a human malignant choroids plexus papilloma cell line (HIBCPP)

A cell line designated “HIBSPP” was established from a human malignant choroids plexus papilloma of 29-year-old Japanese woman. This line grew well without interruption for 3 years and was subcultivated over 70 times. The cells were spindle, oval, and polygonal in shape, and neoplastic and pleomorphic features, a jigsaw puzzle-like arrangement, multilayering and forming papillary shuctures without contact inhibition. The cells proliferated slowly, and the population doubling time was about 69 hours. The chromosome number showed a wide distribution of aneuploidy. The mode was in the hypo-tetraploid range, and many marker chromosomes were observed. The culture cells were easily transplanted into the subcutis of nude mice and produced the tumor resembling the original tumor.

Characterization of newly established human ovarian carcinoma cell line—Special reference of the effects of cis-platinum on cellular proliferation and release of CA125

The cell line HTOA was established from a well-differentiated human ovarian serous cystadenocarcinoma. This line grew well and without interruption for 51 months and was subcultivated over 130 times. The cells were epithelial in shape, and neoplastic and pleomorphic features, a jigsaw puzzle-like arrangement, desmosomal junctional complexes, and multilayering without contact inhibition. The chromosome number was stable at a hypertetraploid range. The culture cells transplanted into BALB/c nude mice and or hamster cheek pouch produced serous cystadenocarcinomas. The cells were found to produce an antigen (CA125) of ovarian cancer, both in vitro and in vivo. The CA125 levels correlated with cellular proliferation in vitro and also with tumor growth, in the nude mouse. These results indicate that the amount of CA125 in the serum is a good marker for detecting early stages of ovarian cancer and in particular for the evaluation of anticancer drugs.

Tumor angiogenic activity of gynecologic tumor cell lines on the chorioallantoic membrane

Tumor angiogenic activity (TAA) from tumor angiogenesis factor (TAF), produced by 24 cell lines of various kinds of gynecologic tumors, was assayed onto chorioallantoic membranes (CAMs) of chick embryos. Methylcellulose (1%) pellets containing 1 x 10(7) cells were placed on 8-day-old postfertilized CAMs, and the grade of neovascularization was assayed 3 days after inoculation. Neovascularization occurred prominently in such cell lines, as HTBOA (poorly differentiated ovarian carcinoma), HUOCA-II (poorly differentiated clear cell adenocarcinoma), HWUA (poorly differentiated endometrial adenocarcinoma), and in HKUS (uterine cervical small cell carcinoma); however, neovascularization did not occur in SNK (uterine leiomyosarcoma line). The cell lines which secreted TAF showed high heterotransplantability in the nude mice and produced rapidly growing tumors which were rich in blood vessels. However, the SKN line which did not secret TAF was not transplantable. These results suggested that there was a close relationship among TAA, transplantability, and tumor growth rate.

Establishment and characterization of two human ovarian endometrioid carcinoma cell lines (with or without squamous cell component)

The cell lines designated HMOA and HNOA were established from human ovarian adenoacanthoma and from a mouse graft of human ovarian endometrioid adenocarcinoma, respectively. These cell lines grew well without interruption for over 20 months. The cultured cells of both HMOA and HNOA lines were spindle, polygonal, and columnar, and showed a jigsaw puzzle-like arrangement and a piling-up tendency devoid of contact inhibition. When the HMOA cells were maintained at the confluent stage, the cells formed cysts and/or squamous metaplasia. The chromosome number of both cell lines varied widely and showed aneuploidy, while the modal chromosome number was stable at the diploid range. Both of these cell lines, HMOA and HNOA, were transplanted into the subcutis of BALB/c nude mice and produced well-differentiated adenoacanthoma and poorly differentiated endometrioid adenocarcinoma, respectively. HMOA cells were characterized as producing large amounts of CA125 (ovarian carcinoma marker), in vitro, in the cyst-forming phase. The HNOA cells, however, did not produce CA125.

Heterotransplantation of mixed mesodermal tumor cells in nude mouse—Histology of metastatic foci

The mixed mesodermal tumor has an admixture of carcinoma and sarcoma. The HIRS-BM tumor cell line, derived from the mixed mesodermal tumor, was transplanted into the peritoneal cavity of BALB/c nude mice, and the metastatic foci were examined histologically and karyologically. The HIRS-BM cells produced mixed mesodermal tumors (admixture of adenocarcinoma and rhabdomyosarcoma) resembling the original tumor. The normal interstitial tissue was not present on the boundary of the sarcoma and carcinoma. These results support the combination theory as the cause of mixed mesodermal tumor.

Establishment and characterization of a human ovarian anaplastic carcinoma cell line

Ovarian carcinoma cell line HTBOA was established from a human ovarian anaplastic carcinoma. It has been proliferating stably since the beginning of culture 3 years ago after 91 passages. The cells were spindle like, oval, and polygonal in shape, and the nuclear chromatin was coarse in grain with many nucleoli, showing neoplasticity and pleomorphism. The cells proliferated rapidly, and the population doubling time was 24-28 hr. The chromosomes showed a wide distribution of aneuploidy, the mode was in the hypotriploid range, and many marker chromosomes were observed. Heterotransplantation was easy, and subcutaneous transplantation of 1 X 10(5) cells in nude mice forms a tumor that is histologically similar to the original anaplastic carcinoma. The most noteworthy characteristic of the cell line was that it caused granulocytosis in nude mice with carcinomas, and formed the colonies of granulocytes in an in vitro culture system. The cell line was considered to be producing a granulocyte colony-stimulating factor.

Histogenesis of carcinosarcoma and Establishment of leiomyosarcoma cell line (HTMMT) derived from human uterine carcinosarcoma

A cell line designated HTMMT was established from the human uterine carcinosarcoma (composed of leiomyosarcoma and adenocarcinoma) of a 66-year-old Japanese woman. The cell line grew well and 83 serial passages were successively done within 24 months. The cell line contained spindle- or fibrous-shaped cells that revealed neoplastic and pleomorphic features, and multipled rapidly without contact inhibition. These cells were characterized as possessing myofibrils. The karyotype exhibited hyperploidy and the chromosome number was ranged from 87 to 100. The cells were transplanted into an immune-depressed hamster cheek pouch or into nude mouse skin, but produced no tumors. We supported the combination theory for the histogenesis of the carcinosarcoma.

Establishment and characterization of a human malignant mesothelioma cell line (HMMME).

A cell line designated “HMMME” was established from the pleural fluids of a malignant mesothelioma patient This line grew well without interruption for 12 years and was subcultured over 200 times. The cells were spindle and roundish in shape and displayed a monolayer sheet in an epithelial pavement cell arrangement. They were neoplastic, had pleomorphic features, and easily formed multilayering without contact inhibition. The cell cytoplasm was strongly positive against anti-vimentin, anti-calretinin and anti-pan-keratin, but negative against anti-BerEP4. The cells proliferated rapidly, and the population doubling time was about 42 hours. Their chromosome number showed a wide distribution of aneuploidy with a mode in the diploid range; many marker chromosomes were observed. The cultured cells were easily transplanted into the subcutaneous of nude mice and produced a tumor classified as a malignant mesothelioma.

Establishment and characterization of a human ovarian endodermal sinus tumor cell line—Producing specific type of α-fetoprotein subfraction

Abstract A cell line was established from an endodermal sinus tumor of the ovary from a 57-year-old Japanese woman, obtained on July 22, 1982. Histological study of the resected tumor revealed a mixture of reticular, solid and cystic patterns, being consistent with the characteristic feature. The cell line, designated as human Akimoto endodermal sinus tumor (HAEST), was subcultivated over 50 times during 29 months. The cells were spindle or columnar in shape and showed pleomorphic and neoplastic features. The modal chromosome number was stable at diploid range and the marker, large acrocentric, chromosome was identified. The cells were transplanted into the cheek pouch of hamsters and produced an endodermal sinus tumor. α-Fetoprotein subfractions in the conditioned media were studied by a modified method of lectin affinity crossed-line immunoelectrophoresis, and we found that the HAEST cells produced concanavalin A nonreactive subfraction, lentil lectin weakly reactive subfraction, phytohemagglutinin-E reactive subfraction and phytohemagglutinin-E nonreactive subfraction, which were predominantly synthesized by the fetal yolk sac, at an early stage of gestation. From these findings, we concluded that the HAEST was indeed an endodermal sinus tumor cell line. This line is expected to have a wide application for various laboratory studies.