• The Journal of Korean Medicine
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• v.30 no.5
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• pp.61-76
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• 2009

Background: Membranous nephropathy (MN) is the most common cause of adult nephrotic syndrome worldwide and has been defined as granular subepithelial deposition of immune complexes along the glomerular basement membrane (GBM). MN has few known treatments and gives rise to side effects under treatment with steroids and immunosuppressives. Objective: The purpose of this experimental study was to demonstrate the effects of Scutellariae Radix extract (SRE) treatment on MN mouse model induced by cBSA. Methods: We divided mice into 4 groups. The Normal group had no treatment. We induced MN mouse model to the other 3 groups by injecting cBSA into the abdominal cavity. The control group was treated with cBSA (10 mg/kg, i.p.) only. The second group, 'SRE-250', was treated with cBSA (10 mg/kg, i.p.) and SRE (250 mg/kg, p.o.). The third group, 'SRE-500', was treated with cBSA (10 mg/kg, i.p.) and SRE (500 mg/kg, p.o.). After cBSA and SRE treatment for 4 weeks, gain in body weight, 24hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN and creatinine of all groups were measured. TNF-$\alpha$, IL-6, IL-1$\beta$, IL-10, IFN-$\gamma$, IgA, IgM and IgG levels of all groups were gauged. H&E staining and electron microscopy of the kidney were observed. Results: SRE showed significant decrease in the 24hrs proteinuria, serum triglyceride, BUN, TNF-$\alpha$, IL-6, serum IgA, IgM and IgG levels compared with the control group. SRE showed increase in the serum IL-10 and IFN-$\gamma$ levels compared with control on RT-PCR. SRE considerably decreased in the thickening of the GBM on H&E staining and deposition of electron-density on electron microscopy of the kidney compared with the control. Conclusions: According to the above results, it is suggested that SRE decreases the symptoms of MN induced by cBSA in mouse model. Therefore, SRE seems to be applicable to MN in clinical practice.

• The Journal of Internal Korean Medicine
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• v.30 no.1
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• pp.212-227
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• 2009

Objective : Membranous nephropathy (MN) is one of the most commonest forms of glomerular disease in man and the most frequent cause of the adult idiopathic nephrotic syndrome. Some investigators recommend no treatment, while others propose aggressive therapy, including prednisolone plus an immunosupressant such as chlorambucil or cyclophosphamide. But a more effective way to treat MN is not defined yet. This study was to evaluate the effects of Patriniae Radix extract (PRE) on the MN induced by cBSA in mice. Methods : Mice were divided into 4 groups. The first group (normal) was injected with saline. The second group (control) was treated with cBSA (10 mg/kg i.p) only. The third group, named PRE-2S0, was treated with cBSA (10 mg/kg i.p) and PRE (250 mg/kg, p.o). The fourth group, PRE-500, was treated with cBSA (10mg/kg i.p) and PRE (500mg/kg, p.o). After cBSA and PRE treatment for 4 weeks, we measured change of body weight, 24hrs proteinuria, serum albumin, total cholesterol, triglyceride, BUN, creatinine, IgG, IgA, IgM, $TNF-\alpha$, $IL-1\beta$, and $IFN-\gamma$ levels and the mRNA expression of $IFN-\gamma$, IL-6, and IL-10. The morphologic changes of renal glomeruli were also observed with a light microscope and an electron microscope. Results : The levels of 24 hrs proteinuria and serum triglyceride. BUN. IgG. $TNF-\alpha$, and $IL-1\beta$ significantly decreased in both PRE groups, while the level of serum albumin significantly increased in both PRE groups. The mRNA expression of IL-10 in splenocytes considerably increased in both PRE groups. The mRNA expression of $IFN-\gamma$ and IL-6 in splenocytes considerably increased in both PRE group. In histological findings of kidney tissue, thickening of GBM decreased in both PRE groups. Conclusions : The present study suggests that PRE is effective when treating mice with MN induced by cBSA. More clinical data and studies are to be done for efficient application.

• Development and Reproduction
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• v.11 no.2
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• pp.85-96
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• 2007

Mesenchymal stem cells (MSC) are promising candidates for cell-based therapies. One major obstacle for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSC. We investigated the effect of human cord serum (HCS) on the growth response, mRNA and protein expressions of human amnion-derived stem cells (HAM). HAM were isolated from the amnion after a Caesarean section and cultured in DMEM supplemented with 10% FBS, 5% HCS or 10% HCS. During culture, their biological characteristics at earlier and later passages were analyzed using RT-PCR and immunocytochemistry. Regardless of serum sources, HAM showed the prominent expression of Oct-4, Rex-1, SCF, FGF-5, BMP-4, nestin, GATA-4, NCAM and HLA ABC genes. The expression profile was observed even at later passages. Similarly, HAM cultured in either FBS or HCS exhibited the distinct protein expression of collagen I, II, III and XII, fibronectin, $\alpha$-smooth muscle actin, vimentin, CK18, CD54, FSP, TRA-1-60, SSEA-3, -4 and HLA ABC. However, desmin expression was only observed in HAM cultured in medium supplemented with FBS and vWF expression was only found in HAM cultured in medium supplemented with HCS. Overall pattern of gene and protein expression of HAM was typical of known adult stem cells such as bone marrow-derived MSC. In conclusion, HCS could be as effective as FBS for the culture of HAM.

• Korean Journal of Veterinary Research
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• v.38 no.3
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• pp.589-594
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• 1998

The in vitro screening tests against the in vitro cultivated $L_3$ of Ascaris suum (in vitro $L_3$), which were cultivated from the embryonated egg to third-stage larva on 7 days in culture(DIC) and the in vivo rat's lung-derived $L_3$ of Ascaris suum (in vivo $L_3$), which were recovered from the lungs of rat on 7 days after infection, carried out in order to compare the anthelmintic efficacy of in vitro $L_3$ and that of in vivo $L_3$ in RPMI medium 1640 with 5% bovine calf serum. And also a screening test of efficacy against adult worms of Trichuris suis performed. The efficacies of screening tests were as follows : 1. The screening efficacies of abamectin and ivermectin against the in vitro $L_3$ were all 100% at the 10ppm concentration in RPMI medium 1640 on 5 DIC. 2. The screening efficacies of abamectin and ivermectin against the in vivo $L_3$ were all 100% at the 20ppm on 5 DIC or at 40ppm on 3 DIC. 3. The screening efficacies of abamectin and ivermectin against the adult worms of Trichuris suis were all 100% at 20ppm on 4 DIC. And therefore, the in vitro cultivated $L_3$ of Ascaris suum were used in the screening test as well as the in vivo rat's lung-derived $L_3$ of Ascaris suum. And also the adult worms such as Trichuris suis and filaroids which is small size and difficult to cultivate to vitro, were used in the screening test in vitro.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2004.10a
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• pp.26-31
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• 2004

This work was undertaken in order to study the developmental competence of nuclear transfer cat embryo with fetal fibroblast and adult skin fibroblast as donor nuclei. Oocytes wererecovered by mincing the ovaries in Hepes-buffered TCM199 and selected the cumulus oocyte complexes (COCs) with compact cumulus cell mass and dark. Homogenous ooplasm were cultured for maturation in TCM199 + 10% fetal bovine serum (FBS) for 12 hours and used as a source of recipient cytoplast for exogenous somatic nuclei. In Experiment 1, we evaluated the effect donor cell types on the reconstruction and development of cloned embryos. Fusion, first cleavage and blastocyst developmental rate was not different between fetal fibroblast and adult skin cell (71.2 vs. 66.8; 71.0 vs. 57.6; 4.0 vs. 6.1 %, P<0.05). In Experiment 2, cloned embryos were surgically transferred into the oviducts of recipient queens. One of seven recipient queens was delivered naturally 2healthy cloned cats and 1 stillborn from fetal fibroblast cell of male origin after 65 days embryo transfer. One of three recipient queens was delivered naturally 1 healthy cloned cat from adult skin cell of female after 65 days embryo transfer. The cloned cats showed genotypes identical to the donor cell lines, indicating that adult somatic cells can be used for feline cloning.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.223-228
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• 2009

We attempted to control the maturation promoting factor (MPF) activity and investigated the subsequent reprogramming of bovine somatic cell nuclear transfer (SCNT) embryos. Serum-starved adult skin fibroblasts were fused to enucleated oocytes treated with 2.5 mM caffeine or $150\;{\mu}M$ roscovitine. The MPF activity, nuclear remodeling patterns, chromosome constitutions and development of SCNT embryos were evaluated. Methylated DNA of embryos was detected at various developmental stages. The MPF activity was increased by caffeine treatment or reduced by roscovitine treatment (p<0.05). Blastocyst development was higher in the caffeine-treated groups (27.6%) than that of the roscovitine-treated group (8.3%, p<0.05). There was no difference in the apoptotic cell index among the three groups. However, the mean cell number of blastocysts was increased in the caffeine-treated group (p<0.05). Higher methylation levels were observed in the Day 3 embryos of the roscovitine-treated group (50.8%), whereas lower methylation levels were noted at Day 5 in the caffeine-treated group (12.5%, p<0.05). These results reveal that the increase in MPF activity via a caffeine-treatment creates a more suitable condition for nuclear reprogramming after SCNT.

• Journal of Biomedical Engineering Research
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• v.19 no.3
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• pp.215-224
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• 1998

This paper is fundamental study to develope the extracorporeal liver support system for patient with fulminant hepatic failure(FHF) or being expected for orthotopic liver transplantation. The polyurethane foam, which is composed of the density of 33kg/m3, the average pore diameter of 500${\mu}{\textrm}{m}$, the closed window of 60-70%, was manufactured with the prepolymer of 15% NCO-, Hepatocytes were inoculated to form spheroids in polyurethane foam. The time of spheroid formation in BSA(Bovine Serum Albumin) coated polyurethane foam was shorter than that in raw polyurethane foam. To verify the function of hepatocyte spheroids, we measured ammonia removal rate, urea and albumin secretion rate. Polyurethane foam was suitable for culture of hepatocyte spheroids. And culture of hepatocyte spheroids in polyurethane foam has high possibility in using as an extracorporeal liver support system.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.140-140
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• 2003

The study evaluated the effect of donor cell treatments for G0/Gl synchronization and the donor ceil type on development and incidence of apoptosis in cloned cattle embryos. Primary cultures were established from a female fetus on day 50 of gestation and adult ear skin biopsies. Cells were randomly allocated into 3 experimental treatment groups after 6~8 passages. Group 1 (Confluent), cells were cultured in DMEM supplemented with 10% FBS until 90% confluent. Group 2 (Serum-starvation), cells were cultured in DMEM Supplemented With 0.5% FBS for 5 days. Group 3 (Roscovitine), Cells were cultured in DMEM supplemented with 10% FBS and 30 $\mu$M Roscovitine for 12 h. Cell cycle and apoptosis were analyzed using flow cytometry after labelling with DAPI and YO-PRO-1. At 19 h post-maturation (hpm), enucleated oocytes were reconstructed with donor cells and fused by a single DC pulse (1.6 kV/cm, 60 $\mu$sec). (중략)

• Clinical and Experimental Pediatrics
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• v.46 no.2
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• pp.167-172
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• 2003

Purpose : Pulmonary vascular hypertension is a common problem in congenital heart disease, the most common cardiac condition in childhood. However, the mechanisms responsible for this pathologic change, treatment, and prevention are poorly understood. Therefore, we studied the gene expression of vascular endothelial growth factor(VEGF) by using a hypoxic model of the pulmonary artery smooth muscle cells. Methods : The main pulmonary artery and its proximal branches of a 6 wk old Fischer rat were excised. They were cut into multiple small pieces and suspended in DMEM medium supplemented with 20% fetal bovine serum and incubated in 5% $CO_2$-95% air atmosphere. The smooth muscle cells were confirmed by immunostaining with smooth muscle myosin and ${\alpha}$-smooth muscle actin antibodies. The VEGF gene expression in the hypoxic group was compared with the one in control the group as well as the one in the starved group by RT-PCR and Northern blot hybridization. Results : There was no statistically significant difference among the control, hypoxic and starved groups. Conclusion : There are few studies of pulmonary vascular hypertension at the molecular level in Korea. Therefore, we studied the expression of VEGF gene in hypoxic pulmonary vascular smooth muscle cells. Further studies will be needed to find the difference between newly born and adult rats, or human and rat pulmonary vascular smooth muscle cells in gene expression. We hope that the study will lead to a better understanding of pulmonary vascular hypertension.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1994.04a
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• pp.131-136
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• 1994

In the present study, in order to make an in vitro model of gastric mucosa for physiological and pharmacological studies, we established two immortalized gastric surface mucous cell lines (GSM06 and GSM10), which produce periodic acid-Schiff (PAS)-and concanavalin A (Con A)-positive glycoproteins, from a primary culture of gastric fundic mucosal cells of adult transgenic mice harboring a temperature-sensitive simian virus 40 large T-antigen gene 〔1]. Gastric fundic mucosal cells were isolated as a modification of a previously described method for rats by Schepp et al. (2). The isolated gastric fundic mucosal cells were cultured in DME/F12 medium supplemented with 2% fetal bovine serum (FBS), 1% ITES (consisting of 2 mg/1 insulin, 2 mgg/1 transferrin, 0.122 mg/1 ethanolamine and 0.00914 mg/1 sodium selenite) and 10 ng/ml recombinant epidermal growth factor (EGF) in a collagen-coated culture dish. To remove fibroblastic cells from the culture, gastric mucosal cells were incubated in the culture medium containing dispase (25 U/ml) for 24 h. The cells, uncontaminated with fibroblastic cells, were then cloned by colony formation. In our series of three attempts, two cell lines (GSM06 and GSM10) have been established at last. The cells proliferated, attached to the dish ana grew until confluent monolayers were formed, and maintained tight contact with neighboring cells. Both GSM06 and GSM10 cells have now been in culture for more than 9 months with regular passaging. The either cell produced