• Biomolecules & Therapeutics
• /
• v.26 no.5
• /
• pp.464-473
• /
• 2018

Cripto is a small glycosylphosphatidylinositol-anchored signaling protein that can detach from the anchored membrane and stimulate proliferation, migration, differentiation, vascularization, and angiogenesis. In the present study, we demonstrated that Cripto positively affected proliferation and survival of mesenchymal stem cells (MSCs) without affecting multipotency. Cripto also increased expression of phosphorylated janus kinase 2 (p-JAK2), phosphorylated signal transducer and activator of transcription 3 (p-STAT3), 78 kDa glucose-regulated protein (GRP78), c-Myc, and cyclin D1. Notably, treatment with an anti-GRP78 antibody blocked these effects. In addition, pretreatment with STAT3 short interfering RNA (siRNA) inhibited the increase in p-JAK2, c-Myc, cyclin D1, and BCL3 levels caused by Cripto and attenuated the pro-survival action of Cripto on MSCs. We also found that incubation with Cripto protected MSCs from apoptosis caused by hypoxia or $H_2O_2$ exposure, and the level of caspase-3 decreased by the Cripto-induced expression of B-cell lymphoma 3-encoded protein (BCL3). These effects were sensitive to down-regulation of BCL3 expression by BCL3 siRNA. Finally, we showed that Cripto enhanced expression levels of vascular endothelial growth factor (VEGF), fibroblast growth factor (FGF), and hepatocyte growth factor (HGF). In summary, our results demonstrated that Cripto activated a novel biochemical cascade that potentiated MSC proliferation and survival. This cascade relied on phosphorylation of JAK2 and STAT3 and was regulated by GRP78. Our findings may facilitate clinical applications of MSCs, as these cells may benefit from positive effects of Cripto on their survival and biological properties.

• Korean Journal of Animal Reproduction
• /
• v.26 no.4
• /
• pp.329-338
• /
• 2002

This study was carried out to investigate the developmental rates of embryo reconstructed with different cell type and to estimate correlation of transcriptional level of octamer-binding transcription factor 4 (Oct4) and fibroblast growth factor 4 (FCF4) gene on peri-implantation stage embryos. Donor cells were transferred into perivitelline space of enucleated oocytes. The karyoplast-cytoplast couplets were accom- plished by cell to cell fusion and activated with ionomycin and 6-dimethylaminopurine. Reconstructed embryos were co-cultured with bovine oviduct epithelial cells in CR 1 aa medium. There is no difference in blastocyst formation rate following nuclear transfer UT) with fetal fibroblast cell (16/50; 32.0%), cumulus cell (16/49; 32.6%) and ear cell (17/52; 32.6%). The expression level of Oct4 and FCF4 in peri-implantation bovine embryo derived from in vitro fertilization (IVF) and NT were determined by reverse-transcription polymerase chain reaction (RT-PCR) technique. In peri-implantation of IVF result in a transient increased of FCF4 paralleled by an increased expression of Oct4. However, Oct4 gene was highly expressed in hatching blastocysts derived from NT compared to IVF. Also, FGF4 expression level in hatching blastocysts and outgrowth stage derived from NT was lower than that of IVF. In conclusion, it is suggested that the different transcription patterns observed in nuclear transfer embryos may lead to a lower rate of embryo development, implantation and pregnancy.

• Journal of Dental Rehabilitation and Applied Science
• /
• v.36 no.3
• /
• pp.145-157
• /
• 2020

Purpose: This study was to determine the possible effects of trichloroacetic acid (TCA) and epidermal growth factor (EGF) through cell survival genes of the PI3K-AKT signaling pathway when applying an hydrophobically modified glycol chitosan (HGC)-based nanocontrolled release system to human gingival fibroblasts in oral soft tissue regeneration. Materials and Methods: An HGC-based nano-controlled release system was produced, followed by the loading of TCA and EGF. The group was divided into control (CON), TCA-loaded nano-controlled release system (EXP1), and the TCA- and EGF- individually loaded nano-controlled release system (EXP2). A total for 29 genes related to the PI3K-AKT signaling pathway were analyzed after 48h of culture in human gingival fibroblasts. Real-time PCR, 1- way ANOVA and multiple regression analysis were performed. Results: Cell survival genes were significantly upregulated in EXP1 and EXP2. From multiple regression analysis, ITGB1 was determined to be the most influential factor for AKT1 expression. Conclusion: The application of TCA and EGF through the HGC-based nano-controlled release system can up-regulate the cell survival pathway.

• Korean Journal of Veterinary Research
• /
• v.52 no.2
• /
• pp.99-104
• /
• 2012

The present study was performed to evaluate the relationship between the neurotoxicity of acrylamide and the differential gene expression pattern in mice. Both locomotor test and rota-rod test showed that the group treated with higher than 30 mg/kg/day of acrylamide caused impaired motor activity in mice. Based on cDNA microarray analysis of mouse brain, myelin basic protein gene, kinesin family member 5B gene, and fibroblast growth factor (FGF) 1 and its receptor genes were down-regulated by acrylamide. The genes are known to be essential for neurofilament synthesis, axonal transport, and neuroprotection, respectively. Interestingly, both FGF 1 and its receptor genes were down-regulated. Genes involved in nucleic acid binding such as AU RNA binding protein/enoyl-coA hydratase, translation initiation factor (TIF) 2 alpha kinase 4, activating transcription factor 2, and U2AF 1 related sequence 1 genes were down-regulated. More interesting finding was that genes of both catalytic and regulatory subunit of protein phosphatases which are important for signal transduction pathways were down-regulated. Here, we propose that acrylamide induces neurotoxicity by regulation of genes associated with neurofilament synthesis, axonal transport, neuro-protection, and signal transduction pathways.

• Proceedings of the Korea Society of Poultry Science Conference
• /
• 2001.11a
• /
• pp.40-46
• /
• 2001

The use of pluripotent stem cells has tremendous advantages for various purposes but these cell lines with proven germ-line transmission have been completely established only in the mouse. Embryonic germ (EG) cell lines are also pluripotent and undifferentiated stem cells established from primordial germ cells (PGCs). This study was conducted to establish and characterize the chicken EG cells derived from gonadal primordial germ cells. We isolated gonadal PGCs from 5.5-day-old (stage 28) White leghorn (WL) embryos and established chicken EG cells lines with EG culture medium supplemented with human stem cell factor (hSCF), murine leukemia inhibitory factor (mLIF), bovine basic fibroblast growth factor (bFGF), human interleukin-11 (hIL-11), and human insulin-like growth factor-I (hIGF-I). These cells grew continuously for 4 months (10 passages) on a feeder layer of mitotically active chicken embryonic fibroblasts. These cells were characterized by screening with the Periodic acid-Shiff＇s reaction, anti-SSEA-1 antibody, and a proliferation assay after several passages. As the results, the chicken EG cells maintained characteristics of undifferentiated stem cells as well as that of gonadal PGCs. When cultured in suspension, the chicken EG cells successfully formed an embryoid body and differentiated into a variety of cell types when re-seeded onto culture dish. The chicken EG cells were injected into blastodermal layer at stage X and dorsal aorta of recipient embryo at stage 14 (incubation of 53hrs) and produced chimeric chickens with various differentiated tissues derived from the EG cells. The germline chimeras were also successfully induced by using EG cells. Thus, Chicken EG cells will be useful for the production of transgenic chickena and for studies of germ cell differentiation and genomic imprinting.

• Journal of Physiology & Pathology in Korean Medicine
• /
• v.25 no.3
• /
• pp.496-502
• /
• 2011

The present study was undertaken to investigate the effect of bee venom acupuncture therapy on the hair follicle growth by macroscopic, microscopic and immunohistochemical observation of skin 10 and 17 days after treatment. The results were as follows : Macroscopic hair follicle growth of 0.2 ml S.B.V. acupuncture treated group was more prominent than those of 0.1 ml S.B.V. acupuncture treated group and control group. Microscopic observation indicated that the hair follicle growth of control group reached anagen phase IV of hair growing cycle, and that of 0.1 ml and 0.2 ml S.B.V. acupuncture treated groups reached anagen phase VI and catagen, respectively. Immunohistochemical observations of the expression of various cytokines, enzymes and receptors in association with hair follicle cycle after local treatment of S.B.V. acupuncture therapy are as follows: Expression of fibroblast growth factor was more intense in epidermis and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of 0.1 ml S.B.V. acupuncture treated group and control group. Expression of epidermal growth factor was more intense in bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of 0.1 ml S.B.V. acupuncture treated group and control group. Expression of c-kit receptor was more intense in epidermis, bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of control group. Expression of vascular endothelial growth factor was more intense in epidermis, bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than that of control group. Expression of protein kinase C-${\alpha}$ was more intense in epidermis, bulge and outer root sheath in 0.2 ml S.B.V. acupuncture treated group than control group. It is concluded that bee venom acupuncture therapy promoted the expression of various cytokines, enzymes and receptors related to the hair growth cycle for hair growth. This findings indicates that bee venom acupuncture therapy is applicable to the treatment of hair loss.

• Journal of the korean academy of Pediatric Dentistry
• /
• v.32 no.3
• /
• pp.395-402
• /
• 2005

In considering the healing process of injured periodontal tissue, healing rate would be influenced by the cellular activity of periodontal fibroblasts(PDLFs). In addition, the reattachment among PDLFs should be induced for healing process. The purpose of this study was to evaluate the effects of epidermal growth factor(EGF) on the proliferation and attachment of PDLFs and to verify the efficacy of EGF as a storage media or a pre-replantation conditioner of traumatically avulsed tooth. Human recombinant epidermal growth factor(hrEGF) and human periodontal fibroblasts from first premolar were prepared. At first, MTT assay was done to evaluate the toxic effect on human periodontal fibroblast and the maximum cellular growth of EGF. Cellular proliferation rate was then compared between control group and 10ng/ml EGF added group. Also, western blot was done to evaluate the expression of fibronectin in both groups. The results were as follows: 1. From MTT assay, EGF showed no toxic effect on PDL fibroblasts. The highest proliferation was shown at 10ng/ml EGF. 2. In 10ng/ml EGF added group, the degree of proliferation of PDLFs was significantly higher than that in control group. 3. Fibronectin expression of EGF added group was also significantly higher than that of control group. From this study we could conclude that EGF enhanced the regeneration rate of periodontal fibroblast, which could be used as a pretreatment agent or a storage media for traumatically avulsed teeth.

• Journal of Applied Biological Chemistry
• /
• v.63 no.1
• /
• pp.75-82
• /
• 2020

In this study, extracts from the Green ball apple peel (GBE) and the newly bred green ball apple from Korea showed inhibition effects on photo-aging factor regulation associated with skin aging. To investigate the inhibition effect on photo-aging factor regulation in skin, GBE was treated with UVB to induce photo-aging related factors in CCD986sk fibroblast cells. Photo-aging factor regulation effects showed that GBE inhibited UVB-stimulated matrix metalloproteinase (MMP)-1 and MMP-9 protein synthesis in collagen type I alpha 2 chain (COL1A2), MMP-1, MMP-9, and tissue inhibitors of metalloproteinase (TIMP)-1 protein expression. The expression of COL1A2 and TIMP-1 protein was significantly increased. The mRNA expression levels of COL1A2, MMP-1, MMP-9, hyaluronan synthase (HAS)2, transforming growth factor (TGF)-β, and TIMP-1 were decreased by GBE. The expression of TIMP-1 and TGF-β, which are regulators involved in matrix metalloproteinase and type I procollagen expression, was found to increase with increasing expression of COL1A2. The expression of HAS2, which is involved in the production of hyaluronic acid, one of the structural proteins constituting the skin, was also confirmed. Therefore, GBE showed excellent efficacy against photo-aging factor regulation and could be used as functional material to prevent and treat skin aging.

• Archives of Plastic Surgery
• /
• v.32 no.5
• /
• pp.625-635
• /
• 2005

In this study, the effects of verapamil on growth rate, apoptosis, production of transforming growth factor (TGF-${\beta}$) and fibronectin were evaluated in keloid and normal human dermal fibroblasts. Both fibroblasts were primarily cultured from earlobe keloids of three female patients and treated with various concentrations of verapamil. Cell toxicity was assessed by MTT assay, growth rate and apoptosis by FACS, and the production of TGF-${\beta}$ and fibronectin by ELISA and Western blot, respectively. In the $MTT_{50}$, the cell growth was more suppressed in keloid fibroblasts. In the $MTT_{90}$, cell growth was more stimulated in normal fibroblasts. No significant effect appeared on TGF-${\beta}$ expression but an increase in extracellular fibronectin secretion was found in keloid fibroblasts. Keloid fibroblasts responded to verapamil more sensitively, and the percentage of apoptosis was higher at the $MTT_{50}$l. In brief, verapamil had growth-inhibitory effect with inducing apoptosis at the $MTT_{50}$, but rather growth-stimulatory effect at the $MTT_{90}$. The biphasic effect of verapamil depending on the dose might explain one of the reasons of relapse after keloid treatment with verapamil. Clinical application with high concentration (2.5 mg/ml) is advised unless excessive dosage is used.

• Animal cells and systems
• /
• v.7 no.3
• /
• pp.221-225
• /
• 2003

The tadpole larva of the ascidian Halocynthia roretzi has two sensory pigment cells in its brain vesicle. To elucidate the temporal requirement for FGF signaling in formation of the pigment cells, embryos were treated with an FGF receptor 1 inhibitor, SU5402, or an MEK inhibitor, U0126 during various embryonic stages. In the present study, it is shown that the embryos treated with SU5402 from the 16-cell stage to the early gastrula stage do not form pigment cells, whereas those treated after the early gastrula stage form pigment cells. In pigment cell formation, embryos suddenly exhibited the sensitivity to SU5402 only for 1 h at the neural plate stage(-4 h after the beginning of gastrulation). When U0126 treatment was carried out at various stages between the 8-cell and late neurula stages, the embryos scarcely formed pigment cells. Pigment cell formation occurred when the embryos were placed in U0126 at early tail bud stage. These results indicate that FGF signaling is involved in pigment cell formation at two separate processes during ascidian embryogenesis, whereas more prolonged period is required for MEK signaling.