• Preventive Nutrition and Food Science
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• v.1 no.2
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• pp.168-173
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• 1996

The effect of methanol soluble fraction(MSF) of kimchi on the proliferating and differentiating activity of normal rat mammary epithelial cells or organoids in culture were studied. Reconstituted basement membrane, Matrigel, supported the growth and development several different multicellular structures from mammary organoids. The five type colonies of multicellular structures, stellate, ductal, webbed, squamous, and lobulo-ductal colonies, were observed in Matrigel culture. In methanol extract groups, webbed colonies were more and squamous colonies were less than control group. and the lobulo-ductal colonies which is known that it formed in well differentiated mammary epoithelial cells were developed more in MSF treated group than control group. These results showed that methanol extract of kimchi affected on the proliferation and differentiation of normal rat mammary epithelial cells cultured in serum free medium condition.

• International Journal of Oral Biology
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• v.36 no.2
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• pp.51-57
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• 2011

Runx2 and Osterix, the transcription factors for osteoblast differentiation, are known as fundamental factors to regulate the development of calcified tissues. However, the biological functions of these factors in the development of the periodontal tissues remain unclear. In this study, we investigated the distribution of Runx2 and Osterix during periodontal tissue development of the mice. Mandibles from 14-day-old mice were prepared for paraffin section. Serial sections of the mandible containing $1^{st}$ molar tooth germs were obtained as a thickness of $7\;{\mu}m$. Some sections were stained with hematoxylin and eosin. Others were used for immunohistochemistry for PCNA, Runx2, and Osterix. Epithelial cells in growing end of Hertwig's epithelial root sheath (HERS) and mesenchymal cells adjacent to the growing end of HERS expressed PCNA. Undifferentiated mesenchymal cells and hard tissue forming cells like cementoblasts and osteoblasts in early stage of differentiation expressed Runx2. Fully differentiated cementoblasts and osteoblasts secreting matrix proteins expressed Osterix. However, the cells terminated the matrix formation did not express Osterix. Periodontal ligament cells expressed Runx2 and Osterix. Pulp cells expressed Runx2 only. These results suggest that Runx2 and Osterix might regulate the differentiation of cementoblasts in the same manner as osteoblasts. Runx2 might participate in the process of cementoblast differentiation in early stage, whether Osterix might regulate the maturation and matrix synthesis of the cells.

• Journal of the Korea Convergence Society
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• v.10 no.4
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• pp.81-89
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• 2019

This study was analyzed about the relationship between culture microenvironment and cell differentiation of HPV 16 E6/E7-transfected immortalized oral keratinocyte(IHOK). By the alteration of culture environment, IHOK-EF and IHOK-EFKGM were obtained, and the modulation of cell properties was observed by cell proliferation assay, immunofluorescence, microarray, and quantitative real-time PCR analysis. IHOK-EF losed the properties of epithelial cells and obtained the properties of mesenchymal cells, and in the result of microarray analysis, genes related to the inhibition of differentiation such as IL6, TWIST1, and ID2 were highly expressed in IHOK-EF. When the culture environment was recovered to initial environment, these changes were recovered partially, presenting the return of genes involved in the inhibition of differentiation such as IL6, and ID2, particularly. This study will contribute to understand adjustment aspect for cell surviving according to the change of culture microenvironment in the study for determining the cell characteristic, and facilitate therapeutic approach for human disease by applying surviving study according to the change of cancer microenvironment.

• International Journal of Stem Cells
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• v.16 no.4
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• pp.394-405
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• 2023

The differentiation of pluripotent stem cells has been used to study disease mechanisms and development. We previously described a method for differentiating human pluripotent stem cells (hPSCs) into salivary gland epithelial progenitors (SGEPs). Here, cystic fibrosis transmembrane conductance regulator (CFTR) knockout hPSCs were differentiated into SGEPs derived from CFTR knockout hESCs (CF-SGEPs) using the same protocol to investigate whether the hPSC-derived SGEPs can model the characteristics of CF. CF-a disease that affects salivary gland (SG) function-is caused by mutations of the CFTR gene. Firstly, we successfully generated CFTR knockout hPSCs with reduced CFTR protein expression using the CRISPR-Cas9 system. After 16 days of differentiation, the protein expression of CFTR decreased in SGEPs derived from CFTR knockout hESCs (CF-SGEPs). RNA-Seq revealed that multiple genes modulating SG development and function were down-regulated, and positive regulators of inflammation were up-regulated in CF-SGEPs, correlating with the salivary phenotype of CF patients. These results demonstrated that CFTR suppression disrupted the differentiation of hPSC-derived SGEPs, which modeled the SG development of CF patients. In summary, this study not only proved that the hPSC-derived SGEPs could serve as manipulable and readily accessible cell models for the study of SG developmental diseases but also opened up new avenues for the study of the CF mechanism.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.92-98
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• 2000

Mesangial cell has several key roles in thee control of glomerular function: it partocipates in the regulation of glomerular filtration rate, macromolecular clearance, and as both a source and target of numerous hormones and autocrines. Many of these insights into mesangial cell function have been obtained by studying mesangial cells in culture. However, no suitble cell lines have established yet. We here reported the immortalization of rat kidney glomeruar mesangial cell by transfection of E6 and E7 genes of human papillomavirus type 16 (HPV-16) via electroporation and lipofection. The reslts showed that only electroporation could transfect the genes to mesangial cells and the transfected cells maintained the viability for longer than 6 months. Fluorescence microscopic observation showed that cellular contractility and phagocytosis, which are the two main phenotypes of mesangial cells with rat glomerular epithelial cells showed that the growth of mesangial dells was suppressed by epithelial cell, but the growth of epithelisl cells was enhanced by mesangial cells. Moreover, Such results may imply that the glomerular cell-cell interaction plays an important role in the regulation of cell proliferation and differentiation.

• Journal of Pharmaceutical Investigation
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• v.40 no.6
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• pp.321-332
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• 2010

Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

• Archives of Pharmacal Research
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• v.20 no.4
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• pp.297-305
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• 1997

A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, $E_2$, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITCPNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.4
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• pp.1881-1886
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• 2016

Background: CA125 is very helpful in treatment monitoring and detection of epithelial ovarian cancer (EOC) recurrence. However there is controversy as to its accuracy and optimal usage. What is the impact of the CA125 levels before primary surgery treatment to the survival of patients? This study aimed to detect any association of preoperative serum levels with prognosis and survival in EOC patients. Materials and Methods: Our cohort comprised EOC patients in Dr. Sardjito Hospital, Yogyakarta, Indonesia, who complied with follow up. To explore the effect of preoperative CA125 levels and other variables on survival Cox's regression models were applied. Results: A total of 90 cases of EOC who had surgery were available for follow up. The level of CA125 proved to be a prognostic factor for overall survival of EOC patients, with an adjusted HR of 4.10 (p = 0.03). Adjuvant chemotherapy was another prognostic factor, 1 - 2 cycles having an adjusted HR of 0.17 (p = 0.04) and 3 - 8 cycles HR 0.39 (p = 0.06). Other factors such as age of patients adjusted HR 1.54 (p = 0.32), moderate differentiation (adjusted HR 1.61, p = 0.51) poor differentiation (adjusted HR 3.41, p = 0.15), and stage of disease (adjusted HR 1.98,p=0.27) were statistically not significant. However, this might have been because the power of the study was low. Conclusions: Preoperative level of CA125 is a prognostic factor for overall survival in EOC patients. The best cut-off for prognostic classification of CA125 serum level is 70 U/ml.

• Archives of Pharmacal Research
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• v.29 no.5
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• pp.424-429
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• 2006

In the present study, a human mammary epithelial cell (HMEC) culture model was developed to evaluate the potential involvement of carrier-mediated transport systems in drug transfer into milk. Trypsin-resistant HMECs were seeded on $Matrigel^{circledR}-coated$ filters to develop monolayers of functionally differentiated HMEC. Expression of the specific function of HMEC monolayers was dependent of the number of trypsin treatments. Among the monolayers with different numbers of treatment (treated 1 to 3 times), the monolayer treated 3 times (3-t-HMEC monolayer) showed the highest maximal transepithelial resistance and expression of $\beta-casein$ mRNA as an index of differentiation. Transport of tetraethylammonium (TEA) across the 3-t-HMEC monolayer in the basolateral-to-apical direction was significantly higher than that in the apical-to-basolateral direction (p<0.05), whereas such directionality was not observed for p-aminohippurate, suggesting the existence of organic cation transporters, but not organic anion transporters. In fact, expression of mRNAs of human organic cation transporter (OCT) 1 and 3 were detected in the 3-t-HMEC monolayer. These results indicate that the 3-t-HMEC monolayer is potentially useful for the evaluation of carrier-mediated secretion of drugs including organic cations into human milk.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.3
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• pp.172-176
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• 2010

Postirradiation extraosseous osteogenic sarcomas are uncommon in the head and neck, despite the extensive use of high-dose radiation. It has been described as de novo radiation-induced neoplasm. We present a 73-year-old male who had been treated by radiotherapy for gingival cancer 7 years earlier and later developed extraosseous osteogenic sarcomas (EOSs) of the neck. Microscopically, the neck mass was composed with mesenchymal malignant cells with cartilaginous and osteogenic differentiation. Immunohistochemical stain demonstrated strong positivity of tumor cells for Snail, the one of major epithelial-mesenchymal transition (EMT) inducer. The E-cadherin expression was scarce, showing inverse relationship to Snail expression. Compared with previous squamous cell carcinoma (SCC) of the gingiva, the present EOS sample revealed the remained epithelial cells on cytokeratin immunohistochemistry, suggesting the tumor arise from the cells of epithelial origin. We have also reviewed the previous 6 cases of head and neck EOSs carefully. The clinicopathologic features of the unusual lesion suggest that it is an incomplete EMT of precedent epithelial malignancy rather than de novo pathology.