[show abstract][hide abstract] ABSTRACT: Atrophy or hypofunction of the salivary gland because of aging or disease causes hyposalivation and has an effect on the quality of life of patients, for example not only dry mouth but deterioration in mastication/deglutition disorder and the status of oral hygiene. Currently conducted therapies for atrophy or hypofunction of the salivary gland in clinical practice are only symptomatic treatments with drugs and artificial saliva, and therefore it is preferable to establish a radical therapy. At this time, as a fundamental investigation, by co-culturing mouse early ES (mEES-6) cells with human salivary gland-derived fibroblasts (hSG-fibro), differentiation of mEES-6 cells to salivary gland cells has been attempted. Also, the possibility of cell engraftment was examined. After identifying the cells which were co-cultured with GFP-transfected mEES-6 cells and hSG-fibro, the cells were transplanted into the submandibular gland of SCID mice, and the degree of differentiation into tissues was examined. The possibility of tissue functional reconstitution from co-cultured cells in a three-dimensional culture system was examined. Our results confirmed that the co-cultured cells expressed salivary gland-related markers and had an ability to generate neo-tissues by transplantation in vivo. Moreover, the cells could reconstitute gland structures in a three-dimensional culture system. By co-culture with hSG-fibro, mEES-6 cells were successfully differentiated into salivary gland cells which were transplantable and have tissue neogenetic ability.
[show abstract][hide abstract] ABSTRACT: The aim of the present study was to isolate endothelial cells from tooth buds (unerupted deciduous teeth) of miniature swine. Mandibular molar tooth buds harvested from swine fetuses at fetal days 90-110 were cultured in growth medium supplemented with 15% fetal bovine serum in 100-mm culture dishes until the primary cells outgrown from the tooth buds reached confluence. A morphologically defined set of pavement-shaped primary cells were picked up manually with filter paper containing trypsin/ethylenediamine tetraacetic acid solution and transferred to a separate dish. A characterization of the cellular characteristics and a functional analysis of the cultured cells at passages 3 to 5 were performed using immunofluorescence, a reverse transcriptase polymerase chain reaction assay, a tube formation assay, and transmission electron microscopy. The isolated cells grew in a pavement arrangement and showed the characteristics of contact inhibition upon reaching confluence. The population doubling time was ~48 h at passage 3. As shown by immunocytostaining and western blotting with specific antibodies, the cells produced the endothelial marker proteins such as vascular endothelial cadherin, von Willebrand factor, and vascular endothelial growth factor receptor-2. Observation with time-lapse images showed that small groups of cells aggregated and adhered to each other to form tube-like structures. Moreover, as revealed through transmission electron microscopy, these adherent cells had formed junctional complexes. These endothelial cells from the tooth buds of miniature swine are available as cell lines for studies on tube formation and use in regenerative medical science.
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] ABSTRACT: This experiment was conducted to address the question of whether acquired paclitaxel resistance is dependent upon whether it is given as a single brief exposure or as a long-term exposure. PX2 and PX24 were established from 2008 human ovarian cancer cells by 2-h single exposure or 24-h continuous exposure to paclitaxel. PX2 acquired paclitaxel resistance faster than PX24 by twofold. Drug resistant pattern was exposure-time dependent. In 2-h exposure, PX2 showed 53.86 ± 4.96 (mean ± standard deviation [SD]) fold paclitaxel resistance while PX24 showed 9.51 ± 1.01 fold resistance (P = 0.002). In 24-h exposure, PX2 showed 2.31 ± 0.3 fold paclitaxel resistance while PX24 showed 28.17 ± 0.98 fold resistance (P = 0.040). PX2 and PX24 acquired cross-resistance to docetaxel and SN38 and the resistance degrees were significantly higher in PX2 than PX24. They displayed approximately twofold cisplatin collateral sensitivity. PX24 also displayed sensitivity to other platinum drugs, oxaliplatin and ZD0473, whereas PX2 acquired significant resistance to both of them. Although differential tubulin-isotype expressions were noted among 2008, PX2 and PX24, they were not significant. In electron microscopy, prominent, densely stained lysosomes were observed more in the resistant cells than 2008. Two independent, exposure-time dependent paclitaxel-resistant human ovarian carcinoma cell lines were established. Understanding the characteristics of the differential resistance pattern could be clinically beneficial for the selection of second line chemotherapy for relapsed ovarian cancer.
Human Cell 11/2010; 23(4):156-63. · 1.41 Impact Factor
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] ABSTRACT: We report a mitochondrial (MT) scoring system related to response to platinum treatment in ovarian cancer (OC). Ultra-thin sections of surgical specimens of primary tumors prepared from 41 OC patients were examined by electron microscopy. The ovarian carcinoma cell line 2008 and its platinum-resistant variant C13 were used as controls. Seven independent MT features, including MT diameter, pattern of cresta structure, electron density, MT distribution, pattern of distribution, ovoid ratio and MT architecture, were examined. Each of the seven parameters was assigned a point score of 0-2 and was summed up with a total score of 14. Clinical response and in vitro sensitivity to platinum, taxane, irinotecan and doxorubicin were evaluated. Clinical information was available for 37 of the 41 cases. Twenty-four cases were stage III and, histologically, 16 serous, 6 endometrioid and 6 clear cell carcinoma were included. All of the patients underwent surgery followed by 6 cycles of taxane and platinum chemotherapy. Fifteen patients exhibited a response, while 22 were resistant to treatment. The total MT score was 5.13±1.13 (mean ± SE) in the 15 responsive cases and 11.41±0.43 in the 22 resistant cases (P<0.001). Receptor operative characteristic (ROC) analysis revealed that the resistant total 'cut-off' score was ≥10 points (P<0.05; AUC=0.86) with 95.5% sensitivity and 80.0% specificity. The MT scoring system correlated well with response to drugs, with the exception of doxorubicin. The progression-free survival (PFS) curves showed an absolute difference in the 6-month PFS of 10% (83 vs. 73%) and in the 12-month PFS of 21% (80 vs. 59%), in favor of patients with low MT scores. This MT scoring system correlates very closely with clinical response as well as cellular sensitivity to chemotherapy, resulting in an association with PFS.
Experimental and therapeutic medicine 01/2010; 1(5):783-788. · 0.34 Impact Factor
[show abstract][hide abstract] ABSTRACT: A cell line designated as NEYS was established from ovarian carcinosarcoma (stage IIIc) of a 56-year-old Japanese woman. The extirpated original tumor was carried in growth medium at 0 degrees C to the culture room. The primary culture was done on 20 August 2003. The cell line was composed of angular adhesive cells and showed neoplastic and pleomorphic features, such as bizarre aggregation of chromatin granules, an irregular thickening nuclear membrane and multiple large nucleoli. They grew as multi-layered cultures without contact inhibition. The cells proliferated moderately, and population doubling time was about 56 h. The chromosome number showed an underdiploidy of aneuploidy. The modal chromosome numbers were 37 (36%) and 38 (26%). The cultures produced carcinoembryonic antigen (27.4 ng/mL), carbohydrate antigen 19-9 (210 U/mL), and carbohydrate antigen 125 (526 U/mL). The NEYS cells did not give rise to transplant tumors in nude mice, and showed no susceptibility against cisplatin (CDDP), CPT-11, carboplatin, Paclitaxel, Taxotere and 5-FU. This cell line is useful for studies on the histogenesis of carcinosarcoma and susceptibility of cancer drugs in human ovarian carcinosarcoma. The immunohistochemical and ultrastructual analysis demonstrated that NEYS cells showed epithelial and mesenchymal differentiation, and supported the metaplasis theory as the cause of carcinosarcoma.
Human Cell 09/2009; 22(3):72-80. · 1.41 Impact Factor
[show abstract][hide abstract] ABSTRACT: Apart from typical yolk sac tumors, ovarian tumors with elevated alfa-fetoprotein (AFP) are uncommon and the differential diagnosis needs to consider the hepatoid pattern of a yolk sac tumor, hepatocellular carcinoma metastatic to the ovary, hepatoid carcinoma, and other epithelial ovarian tumors. We report here an AFP-producing ovarian tumor with uncertain pathological diagnosis, which was extremely responsive to chemotherapy. A 59-year-old Japanese woman presented with lower abdominal distension and was found to have a left ovarian mass on pelvic examination and magnetic resonance imaging (MRI) scan. Laboratory tests showed serum AFP, 73 687 ng/ml; carbohydrate antigen 125 (CA125), 1599 U/ml; and carcinoembryonic antigen (CEA), 13.9 ng/ml. Total hysterectomy with bilateral salpingo-oophorectomy, partial omentectomy, and low anterior resection of the rectum was performed, without any residual macroscopic tumor. Microscopically, the tumor was characterized by a hepatoid carcinomatous component composed of solid sheets of large eosinophilic cells with pleomorphic nuclei. The pathological stage was pT2N0M0. Tumor cells were diffusely immunoreactive for AFP and cytokeratin (CAM5.2), but monoclonal CEA and CA19-9 were focally positive in the cytoplasm, while CA125 was negative. The patient was treated postoperatively with three cycles of chemotherapy consisting of bleomycin, etoposide, and cisplatin; with this regimen, serum AFP decreased to 16 ng/ml from 12 600 ng/ml just before the initiation of chemotherapy. The patient received secondary cytoreductive surgery of systemic lymphadenectomy, which revealed no evidence of residual tumor.
International Journal of Clinical Oncology 03/2009; 14(1):70-3. · 1.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: We developed mitochondrial (MT) scoring system based on MT ultrastructural findings in association with response to chemotherapy in ovarian cancer (OC). Ultrathin sections of MT prepared from 28 OC patients before chemotherapy were examined by electron microscopy. Platinum-sensitive ovarian carcinoma cell line 2008 and its resistant variant C13 were used as control cells. Seven independent MT features including, diameter, pattern of cresta structure, electron density, distribution-density and -pattern, ratio of minimal/maximal diameter and MT architecture were examined and were assigned a score between 0 and 2. Twenty-eight cases were primary advanced OC, including 4 recurrent cases. Nine cases were chemosensitive while 19 were resistant. Univariate and multivariate analysis in each factor showed good correlation to chemosensitivity for 2 factors of electron density, distribution pattern. Total score of these 2 factors in 9 sensitive cells was 1.44+/-0.41 (M +/- SE) and was 3.58+/-0.18 in 19 resistant cells (P<0.001). Receptor operative characteristics (ROC) analysis revealed that total 'cut-off' score was 3 point (P<0.05; AUC=0.84). In conclusion, this MT scoring system was excellently correlated to response regardless of histopathologic findings and this strongly suggests that the system is deemed to be of great value as biomarker for the chemosensitivity in OC.
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] ABSTRACT: Abstract 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 lime 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.
Human Cell 10/2008; 16(4):231 - 239. · 1.41 Impact Factor
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] 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. · 1.22 Impact Factor
[show abstract][hide abstract] 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. · 1.34 Impact Factor
[show abstract][hide abstract] 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. · 1.41 Impact Factor
[show abstract][hide abstract] 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. · 1.41 Impact Factor