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Kazushige Kiguchi,
Isamu Ishwata,
Yuko Tokieda,
Megumi Iguchi,
Chieko Ishiwata,
Masanori Iwata,
Bunpei Ishizuka,
Hiroyuki Yoshikawa,
Toshiaki Tachibana, Hisashi Hashimoto,
Hiroshi Ishikawa
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ABSTRACT: A cell line designated HUUCLEC was established from a human uterine cervical lymphoepithelial carcinoma obtained from a 61-year-old
Japanese woman. The cell line has grown slowly without interruption and serial passages were successively carried out 60 times
within 3 years. The cultured cells were spindle or round in shape, showing anaplastic and pleomorphic features, a pavement
cell arrangement and multilayering without contact inhibition. The population doubling time of the HUUCLEC line was 72 hours
while the chromosomal number varied widely and showed aneuploidy. The modal chromosomal number was stable at the triploid
range and marker chromosomes were present the Ebstein-Barr virus was absent in the cultured cells.
Key wordsCell line–Lymphoepithelioma-like carcinoma–uterine cervix
Human Cell 04/2012; 15(2):97-102. · 1.27 Impact Factor
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ABSTRACT: Currently there is no good hepatocyte model for studying growth hormone (GH) function that reflects its normal physiological roles. Here we report the establishment of a functional hepatocyte cell line, SDRL-1, from the liver of young male spontaneous dwarf rats (SDR), with isolated GH deficiency. This line has been maintained in Dulbecco's Modified Eagle Medium (DMEM)/F12 medium supplemented with 10% fetal bovine serum (FBS) with retention of a near diploid karyotype for extended periods of time. When grown as a monolayer sheet, it displayed a pavement-like appearance and contact inhibition. These cells have a poorly developed rough endoplasmic reticulum (r-ER), few mitochondria and glycogen granules, and produce a small amount of albumin and α-fetoprotein, that is enhanced when grown on a collagen gel sponge. Human recombinant GH stimulated JAK2 and STAT5b tyrosine phosphorylation and IGF-I production in a concentration-dependent manner. When the cells were cultured with GH-supplemented medium, the number of mitochondria and glycogen granules increased together with the r-ER and Golgi apparatus. A number of microvilli were observed on the surface of the cultured cells, further suggesting that this cell line is composed of normally functioning hepatocytes. In summary, we established a novel hepatocyte cell line (SDRL-1), that appears to display normal function, which we propose can serve as a good in vitro model for studying GH-target organ interactions.
Human Cell 11/2010; 23(4):164-72. · 1.27 Impact Factor
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ABSTRACT: There are few case reports describing small cell lung carcinoma (SCLC), which secrete parathyroid hormone (PTH)-related protein (PTHrP) and result in hypercalcemia. We have established a novel cell line, derived from a 37-year-old woman with SCLC, which produced PTH, PTH-rP, and a part of proopiomelanocortin (POMC), and led to hypercalcemia. The cell line, named SS-1, was grown as floating cell clusters in DMEM/F12 medium supplemented with 10% fetal bovine serum and had a population doubling time of 72 h. The modal chromosome number was 47 (88%); marker chromosomes were not observed. The SS-1 cell line secreted not only PTHrP but also PTH, and both were decreased by CaCl(2) administration. Decreasing the concentration of Ca(++) in the growth medium stimulated the secretion of both PTHrP and PTH. The cell line had calcium sensing receptor (Cas-R). Since PTHrP and PTH secretion from the SS-1 cells was related to Ca(++) concentration in the growth medium, the cell line might be useful for the study of PTH-rP and PTH regulation as well as for SCLC analysis. In addition, the cells secreted N terminal POMC, the precursor of adrenocorticotropic hormone, in response to stimulation with corticotropin releasing hormone. In summary, we established a novel cell line, SS-1 from SCLC, which produced PTHrP, PTH and N terminal POMC.
Human Cell 05/2010; 23(2):58-64. · 1.27 Impact Factor
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ABSTRACT: 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-erbB/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 CA15-3, NCC-ST-439 and HER2 protein. We believe the cell line will contribute to the therapeutic study of malignant breast cancer.
Human Cell 12/2008; 21(4):105-12. · 1.27 Impact Factor
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Satoshi OHI,
Shigeki NIIMI,
Naoya OKADA,
Kyosuke YAMADA,
Toshiaki TACHIBANA, Hisashi HASHIMOTO,
Masako NAKAJIMA,
Mitsuru YASUDA,
Tadao TANAKA,
Kahei SATO,
Hiroshi ISHIKAWA
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ABSTRACT: Abstract We successfully established a novel cell line (OS-1) derived from human ovarian small cell carcinoma, hypercalcemic type secreted PTH, PTH-rP and ACTH. The OS-1 cell line was established from metastatic focus of uterus. A patient was 25-year-old Japanese woman. The first she received left ovariectomy on April 2002. The histopathological diagnosis was ovarian small cell carcinoma, pT2c, Nx, Mx. Then on June 2003, metastatic focus of uterus was ectomied. A part of the recurrent tumor of uterus was cut into small pieces with razor blades, and dissociated with 0.1% trypsin-0.02% EDTA/ PBS(−) solution at room temperature. The single cells and small cell clusters were seeded into 60mm dishes and cultured in growth medium (GM: DMEM/F12 supplemented with 20% fetal bovine serum and 0.1% non-essential amino acids solution) at 37°, 4.7% CO2 in humidified air. Medium was exchanged twice a week. The OS-1 cells grew as floating cultures in the dishes. Radioimmunoassay of the conditioned media was revealed that the cultures secreted large amount of PTH, PTHrP and ACTH simultaneously. Susceptibilities of anti-cancer drugs to the OS-1 cells were examined using oxygen electrode meter (Daikin), and the results suggested VLB and TXL were effective, and CDDP, CPT-11, VP-16, VCR, CPA, MMC and CBDCA were not effective.In our knowledge, it is the first report that the cell line secreting PTH, PTHrP and ACTH was successfully established from ovarian small cell carcinoma, hypercalcemic type. We expect that OS-1 cell line contribute to study on the mechanism of ectopic hormone secretion and susceptibility of anti cancer drugs to the small cell carcinoma.
Human Cell 10/2008; 17(4):203 - 210. · 1.27 Impact Factor
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ABSTRACT: Abstract Human gestational choriocarcinoma cell line (HOCC) was established from the mouse graft of choriocarcinoma. The HOCC cells were spindle or polygonal in shape and multi-nucleated giant cells, showing neoplastic and pleomorphic features. The cell proliferated stably, and the population doubling time was about 32 hours. The chromosome numbers showed a wide distribution of aneuploidy, the mode was in hypertriploid range and the marker chromosomes were recognized in the several generations. Heterotransplantation was easy, and subcutaneous transplantation of 1 × 107 cells in nude mouse formed a tumor composed of choriocarcinoma. It is most noteworthy characteristic of the cell line that it produced human chorionic gonadotropin (hCG) in an in vitro culture system and in vivo in nude mice.
Human Cell 10/2008; 17(1):33 - 42. · 1.27 Impact Factor
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ABSTRACT: Abstract Two cell lines, HYVC and HMVC, were established from cultures of the squamous cell carcinoma removed from the vulva of females of 37 and 46 years old, respectively. These cell lines were very similar in their biological characteristics, such as morphology (polygonal), growth pattern (32–43 hr of population doubling time, 50–2596 of plating efficiency), heterotransplantability (1X107 cells produced squamous cell carcinomas in the nude mice), genetics (75–78 of modal chromosomal number), and producing the tumor markers (SCC and TPA). The HPV-DNA was not detected in either the HYVC or HMVC lines by using L1-PCR methods, however, it was detected in the HYVC using E6/E7-PCR.
Human Cell 10/2008; 15(4):207 - 214. · 1.27 Impact Factor
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ABSTRACT: Abstract Angiogenesis is indispensable to guide a regeneration of good periodontal tissue in the wound healing after periodontal surgery. Hepatocyte growth factor is well known for a strong angiogenic factor and it may play important roles in the periodontal tissue during periodontal wound healing. In exploring the promotion of angiogenesis in the periodontal ligament, proliferauve and tubulogenic responses of endothelial cells to hepatocyte growth factor and to soluble factors secreted by fibroblasts were investigated. Pavement-shaped cells isolated from a human periodontal ligament were identified as the endothelial cell by their granular immunoreactivity for factor VIII. The proliferation of the endothelial cells was accelerated by the addition of hepatocyte growth factor or fibroblast-conditioned medium, and far more by adding both than either. The endothelial cells seeded on the agar containing both hepatocyte growth factor and fibroblast products formed a dense network in a shorter time than on the agar containing either. The endothelial cells in the dense network took a tube-like structure with lumen and were covered with laminin. These results suggest that hepatocyte growth factor administered into the regenerating periodontal tissue may promote, synergistically with local factors produced by the activated fibroblast, the proliferation and tubulogenesis of the remaining endothelial cells.
Human Cell 10/2008; 18(2):83 - 91. · 1.27 Impact Factor
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ABSTRACT: Abstract 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 hypotetraploid 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.
Human Cell 10/2008; 18(1):67 - 72. · 1.27 Impact Factor
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ABSTRACT: Cancer cells have characteristics, such as high telomerase activity and high levels of migration activity and proliferation, which are very similar to those of germ cell lineages. In this study, we examined the expression of VASA, a germ cell lineage specific marker and evaluated its clinical significance in epithelial ovarian cancer (EOC).
We investigated VASA expression in 75 EOC tissues by immunohistochemistry, correlating results with clinicopathological factors. To clarify the effects of VASA on cellular phenotypes, we compared the protein expression profiles between SKOV-3 cells stably expressing VASA (SKOV-3-VASA) and vector-control cell lines by coupling 2D fingerprinting and identification of proteins by mass spectrometry.
VASA expression in tumor cells was found in 21 of 75 cases and was positively correlated with high age and serous histology. Significant down-regulation of 14-3-3sigma was observed in SKOV-3-VASA versus control cells. Over-expression of VASA abrogates the G2 checkpoint, induced by DNA damage, by down-regulating the expression of 14-3-3sigma.
These results suggest that VASA may either play a direct role in the progression of EOC or serve as a valuable marker of tumorigenesis.
Gynecologic Oncology 10/2008; 111(2):312-9. · 3.89 Impact Factor
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ABSTRACT: Uterine papillary serous carcinoma is an uncommon histologic subtype of endometrial cancer that behaves aggressively and has a poor prognosis. We successfully established a uterine papillary serous carcinoma cell line. The population-doubling time was approximately 16 h. Although loss of p53 function is considered critical for the molecular pathogenesis of uterine papillary serous carcinoma, p53 was not only mutated but functionally active in this cell line. This newly established cell line should be useful for investigating the characteristics of uterine papillary serous carcinoma.
Human Cell 09/2008; 21(3):64-9. · 1.27 Impact Factor
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ABSTRACT: We recently established human chorionic gonadotropin-, adrenocorticotropic hormone- and parathyroid hormone-related protein-secreting cell line derived from primary poorly differentiated adenocarcinoma of the stomach. The cell line was designated as IGSK-3. Inverted-phase contrast microscopy revealed that the IGSK-3 cells consist of two morphological subtypes. One type has visible nucleoli and clear nuclei, but nucleoli and nuclear membrane of the other type are invisible. The population-doubling time was about 43 h. An analysis of conditioned medium by IGSK-3 cells cultured for 4 days revealed the IGSK-3 cells secrete human chorionic gonadotropin-beta (0.5 ng/mL), adrenocorticotropic hormone (5.5 pg/mL), parathyroid hormone-related protein (3.4 pmol/mL) and epidermal growth factor (14.2 pg/mL). Histopathological diagnosis of the graft of IGSK-3 cells revealed that IGSK-3 cells built a poorly differentiated adenocarcinoma which resembled the original tumor. In addition, the IGSK-3 cell line was immunocytochemically positive for human chorionic gonadotropin-beta and epidermal growth factor receptor, and negative for vascular endothelial growth factor.
Human Cell 09/2008; 21(3):88-94. · 1.27 Impact Factor
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ABSTRACT: Phototherapy is the most frequently used treatment for the neonatal jaundice. However, recent papers report that phototherapy increased apoptosis in peripheral mononuclear leukocytes in vivo and in mouse lymphoma cell line in vitro. We have investigated the cytotoxicity of phototherapy on the small intestine of neonatal rat using conventional halogen-quartz device (conventional device) and blue light-emitting device (LED device) by measuring apoptotic cells. Four-day-old male Wistar rats were divided into three groups as follows: group 1, exposure to conventional device for 72 h; group 2, exposure to LED device for 72 h; and group 3, control (without phototherapy). After light exposure, the small intestine was examined for apoptosis. Apoptotic cells were detected by the TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) assay, by immunohistochemistry for caspase-3 and by transmission electron microscopy. The proportion of positive cells by the TUNEL method in the epithelium of the small intestine was 6.2, 3.1 and 1.7% in the conventional device group, the LED device group and the control group, respectively. The apoptotic cells of the conventional device group is significantly higher than the LED device group (P < 0.01) and that of the LED device group was higher than that of the control group (P < 0.05). We suspected that phototherapy induced apoptosis in neonatal small intestine and the conventional device introduces more apoptosis than the LED device.
Pediatric Surgery International 08/2008; 24(7):837-42. · 1.25 Impact Factor
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ABSTRACT: The formation of the crypt in the distal colon of the mouse was investigated in association with the development of vascular networks. For histological observation, 1-microm cross-sections were made from the distal colon of fetal mice in 13 to 18 days of gestation. Three-dimensional distributions of vascular networks in the organ were observed after perfusing fetuses with rhodamine isothiocyanate-labeled gelatin and immunostaining for laminin to examine the boundary between the epithelium and the mesenchyme. At 13 days of gestation, the distal colon and its epithelium formed a cylindrical tube and a loose primary plexus of vessels was built in the mesenchyme. In the distal colon of 15 days of gestation, the caudal portion began to form the crypt and the vascular plexus built up from a few layers was situated apart from the boundary between the epithelium and the mesenchyme. As the development proceeded, the formation of the crypt occurred in the caudorostral direction. The developing crypt advanced into the vascular plexus, so that a few vessels situated in the mesenchyme between crypts. As the crypt elongated, these vessels formed a small plexus situated perpendicular to the primary plexus, while the primary plexus became monolayered and loosened. The new plexus was composed of ascending vessels and traversing ones, but the regular honeycomb-like plexuses around openings of crypts have not established yet even in 18 days of gestation. The vascular system as well as the crypt in the distal colon will take further a few postnatal weeks to be completed.
The Anatomical Record Advances in Integrative Anatomy and Evolutionary Biology 02/2008; 291(1):65-73. · 1.47 Impact Factor
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ABSTRACT: A novel method for acquiring serial images suitable for three-dimensional reconstruction of vascular networks in the whole brain of mouse was developed. The brain infused with a White India ink-gelatin solution was fixed and embedded in paraffin containing Sudan Black B through xylene also containing Sudan Black B. Each sliced surface of the paraffin block was coated with liquid paraffin and its image was serially acquired. Coating with liquid paraffin extremely improved the quality of the image. The series of serial images was free of distortion and a three-dimensional image was reconstructed without the problem of the alignment and registration of adjacent images. The volume-rendered image indicated three-dimensional distribution of blood vessels in a whole brain. No ghost or shadow was observed on a volume-rendered image of the White India ink-gelatin infused brain. The z-axial resolution examined on the orthogonal sections reconstituted from serial images obtained at an interval of 5 mum showed no cross talk, indicating that the z-axial resolution was no larger than 5 mum. A proper understanding of the vascular system in a whole brain is indispensable to reveal the development of the vascular system in the brain of normal and genetically manipulated mouse and vascular alterations in pathological situation, such as stroke and neurodegenerative disease. Although simple and inexpensive, this method will provide fundamental information on the vascular system in a whole brain.
Microscopy Research and Technique 02/2008; 71(1):51-9. · 1.79 Impact Factor
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ABSTRACT: 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.
Human Cell 09/2007; 20(3):85-90. · 1.27 Impact Factor
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ABSTRACT: We examined a 32-year-old Japanese man who was clinically diagnosed with gastric cancer, type 4, and histopathologically diagnosed with mucinous and poorly differentiated adenocarcinoma (mucinous > poorly) of the stomach. We successfully established and characterized a cell line (designated as IGSK-2) derived from the ascitic fluid of the patient with recurrent and cisplatin-resistant carcinoma. The IGSK-2 cells grew in multi-layered culture in culture dishes. The cells secreted 18 pg/mL somatostatin, 9.1 mIU/mL human chorionic gonadotrophin (hCG), 8000 U/mL carbohydrate antigen 19-9 and 410 ng/mL carcinoembryonic antigen over 4 days of culture. The population doubling time was approximately 83 h. The susceptibility test of anticancer drugs revealed that IGSK-2 cells were sensitive to Taxol, but were not sensitive to cisplatin, 5-fluorouracil and irinotecan. Immunohistochemical staining revealed that the IGSK-2 cells were positive against antihCG antibody and antiserotonin antibody, and negative against antisomatostatin antibody and antigastrin antibody.
Human Cell 06/2007; 20(2):52-61. · 1.27 Impact Factor
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ABSTRACT: We successfully established a spontaneously cisplatin-resistant tumor cell line (designated as IGSK-1) derived from original gastric carcinoma. The patient was a 75-year-old Japanese woman. The histopathological diagnosis was gastric poorly differentiated adenocarcinoma accompanied with metastatic foci in lymph nodes, pT3, N2 M0, stage IIIB. The IGSK-1 cells grew as adhesive and monolayered cultures on the bottom of dishes. The susceptibility of the IGSK-1 cells to anti-cancer drugs was examined using oxygen electrode apparatus (Daikin, Tsukuba, JPN), and the results suggested TXL was effective, and CDDP, CPT-11 and 5-FU were not effective. Gastrin and somatostatin secretions were confirmed by immunohistochemical staining and also radioimmunoassay. Immunohistochemistry and radioimmunoassay for serotonin suggested the IGSK-1 cells might incorporate serotonin from the growth media. Spontaneously cisplatin-resistant gastric carcinoma cell line secreted gastrin and somatostatin is very important material for chemotherapy.
Human Cell 05/2007; 20(1):15 - 22. · 1.27 Impact Factor
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ABSTRACT: We successfully established a spontaneously cisplatin-resistant tumor cell line (designated as IGSK-1) derived from original gastric carcinoma. The patient was a 75-year-old Japanese woman. The histopathological diagnosis was gastric poorly differentiated adenocarcinoma accompanied with metastatic foci in lymph nodes, pT3, N2 M0, stage IIIB. The IGSK-1 cells grew as adhesive and monolayered cultures on the bottom of dishes. The susceptibility of the IGSK-1 cells to anti-cancer drugs was examined using oxygen electrode apparatus (Daikin, Tsukuba, JPN), and the results suggested TXL was effective, and CDDP, CPT-11 and 5-FU were not effective. Gastrin and somatostatin secretions were confirmed by immunohistochemical staining and also radioimmunoassay. Immunohistochemistry and radioimmunoassay for serotonin suggested the IGSK-1 cells might incorporate serotonin from the growth media. Spontaneously cisplatin-resistant gastric carcinoma cell line secreted gastrin and somatostatin is very important material for chemotherapy.
Human Cell 03/2007; 20(1):15-22. · 1.27 Impact Factor
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ABSTRACT: We successfully established a breast scirrhous carcinoma cell line (designated as NABCA) derived from metastatic tumors of the lymph node. The cells grew as multi-layered cultures without contact inhibition. The population doubling time was approximately 66 h. G-band karyotype of NABCA revealed 66% diploid, XX. Surprisingly, the cells had a number of secretory granules and straight microvilli as a brash border. In heterotransplantation, the cells produced a tumor resembling the original tumor. The NABCA is sensitive to Adriamycin (doxorubicin; KYOWA HAKKO KOGYO, Tokyo, Japan) and Taxol (paclitaxel; Bristol-Myers KK, Tokyo, Japan). This cell line is useful for studying the mechanism of lymphatic metastasis and susceptibility of anticancer drugs in human breast scirrhous cancer.
Human Cell 12/2006; 19(4):126-32. · 1.27 Impact Factor
