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Jae-Sung Ryu,
Kyu-Tae Chang,
Ju-Taek Lee,
Malg-Um Lim,
Hyun-Ki Min,
Yoon-Ju Na,
Su-Bin Lee,
Gislain Moussavou,
Sun-Uk Kim,
Ji-Su Kim,
Kinarm Ko,
Kisung Ko,
Kyung-A Hwang,
Eun-Jeong Jeong,
Jeong-Woong Lee, Young-Kug Choo
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ABSTRACT: Gangliosides play important roles in the control of several biological processes, including proliferation and transmembrane signaling. In this study, we demonstrate the effect of ganglioside GM1 on the proliferation of mouse induced pluripotent stem cells (miPSCs). The proliferation rate of miPSCs was lower than in mouse embryonic stem cells (mESCs). Fluorescence activated cell sorting analysis showed that the percentage of cells in the G2/M phase in miPSCs was lower than that in mESCs. GM1 was expressed in mESCs, but not miPSCs. To confirm the role of GM1 in miPSC proliferation, miPSCs were treated with GM1. GM1-treated miPSCs exhibited increased cell proliferation and a larger number of cells in the G2/M phase. Furthermore, phosphorylation of mitogen-activated protein kinases was increased in GM1- treated miPSCs. [BMB Reports 2012; 45(12): 713-718].
BMB reports 12/2012; 45(12):713-8. · 1.72 Impact Factor
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Jin-Hyoung Cho,
Ji-Su Kim,
Malg-Um Lim,
Hyun-Ki Min,
Dong-Hoon Kwak,
Jae-Sung Ryu,
Ju-Taek Lee,
Sun-Uk Kim,
Chang-Hwan Kim,
Chang-Hyun Kim,
Deog-Bon Koo,
Kyu-Tae Chang, Young-Kug Choo
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ABSTRACT: Gangliosides are ubiquitous components of the membranes of mammalian cells that are thought to play important roles in various cell functions such as cell-cell interaction, cell adhesion, cell differentiation, growth control, and signaling. However, the role that gangliosides play in the immune rejection response after xenotransplantation is not yet clearly understood. In this study, the regulatory effects of human leukocytes on ganglioside expression in primary cultured micro-pig aortic endothelial cells (PAECs) were investigated. To determine the impact of human leukocytes on the expression of gangliosides in PAECs, we performed high-performance thin layer chromatography (HPTLC) in PAECs incubated with FBS, FBS containing human leukocytes, human serum containing human leukocytes, and FBS containing TNF-α. Both HPTLC and immunohistochemistry analyses revealed that PAECs incubated with FBS predominantly express the gangliosides GM3, GM1, and GD3. However, the expression of GM1 significantly decreased in PAECs incubated for 5 h with TNF-α (10 ng/mL), 10% human serum containing human leukocytes, and 10% FBS containing human leukocytes. Taken together, these results suggest that human leukocytes induced changes in the expression profile of ganglioside GM1 similar to those seen upon treatment of PAECs with TNF-α. This finding may be relevant for designing future therapeutic strategies intended to prolong xenograft survival.
Laboratory animal research. 12/2012; 28(4):255-63.
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ABSTRACT: To investigate neuroprotective effects of three major anthocyanins (cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, and petunidin-3-O-glucoside) isolated from the black soybean (Glycine max L.) cv. Cheongja 3 seed coat against H(2)O(2)-induced cell death of human brain neuroblastoma SK-N-SH cells.
Cell viability, reactive oxygen species (ROS) generation, production and expression of heme oxygenase (HO)-1 and inactivation of mitogen-activated protein (MAP) kinase cascades were determined by MTT assay, 2,7-dichlorofluorescein diacetate (DCF-DA) assay, reverse transcriptase polymerase chain reaction (RT-PCR), and western blotting, respectively.
Pretreatment with anthocyanins reduced the cytotoxicity of H(2)O(2) on SK-N-SH cells, dose-dependently reduced the intracellular ROS level and inactivated apoptosis signal-regulating kinase (ASK1, Thr845), p38, and c-Jun N-terminal kinase (JNK) proteins. The HO-1 and Neu1 mRNA levels were increased by H(2)O(2) (25 μM) and further elevated by the pretreatment with anthocyanins. Sialic acids added to the culture plates not only attenuated the cytotoxicity of H(2)O(2) (25 μM) but also reduced intracellular ROS level. These results suggest that Cheongja 3 black soybean seed coat anthocyanins have brain neuroprotective effects against oxidative stress (H(2)O(2)) by inhibiting the activation of ASK1-JNK/p38 pathways, scavenging ROS, stimulating the expression of HO-1 and, more interestingly, recruiting cellular free sialic acids through up-regulation of Neu1 sialidase gene expression.
This is the first report indicating potent health benefits of black soybean seed coat anthocyanins in neuroprotection by triggering mobilization of cellular free sialic acid and utilizing it as an additional biological antioxidant in brain neural cells.
Life sciences 04/2012; 90(21-22):874-82. · 2.56 Impact Factor
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ABSTRACT: Baculovirus has been widely used for the production of numerous recombinant proteins in insect cells. Baculovirus vectors
have several advantages, including proper post-translational modification, biosafety, and multiple large gene expression ability.
Most insect cell-produced proteins have been expressed by using the baculovirus expression vector system (BEVS) under the
control of strong polyhedrin (Polh) or p10 promoters. There has been no report on the expression of recombinant proteins by
baculovirus in plant cells. In this study, we used the baculovirus vector to express recombinant green fluorescent protein
(GFP) in plants. To investigate the expression of GFP protein by baculovirus in plants, we cloned the gfp gene under the control
of Polh promoter or Cauliflower Mosaic Virus (CaMV) 35S promoter to yield the Polh-GFP and 35S- GFP bacmids carrying the GFP
expression cassettes, respectively. The presence of Polh-GFP and 35S-GFP expression cassettes in the bacmids was confirmed
by polymerase chain reaction (PCR). Subsequently, both the GFP bacmids and GFP baculovirus vectors generated from the bacmid-transfected
Sf9 insect cells were inoculated into Nicotiana benthamiana leaves. Confocal microscopy revealed that the gfp gene expression was high in plant leaves at 48 and 72 h after bacmid and
baculovirus inoculation. Reverse transcription-PCR (RT-PCR) and fluorescence microscopy confirmed that the gfp genes under
the control of Polh or CaMV35S promoters were highly expressed in plant leaves inoculated with 40 L of baculovirus solution.
These results suggested that the baculovirus vector can be used to express recombinant proteins in plants. The baculovirus
vector- mediated gene delivery and expression system could be used in plant biotechnology for fast and efficient production
of recombinant proteins and for molecular virology studies in plants.
Additional key wordsCaMV 35S promoter–GFP protein–Sf9 insect cell–polyhedrin promoter
04/2012; 52(1):95-104.
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Bong-Seok Song,
Seung-Bin Yoon,
Ji-Su Kim,
Bo-Woong Sim,
Young-Hyun Kim,
Jae-Jin Cha,
Seon-A Choi,
Hyun-Ki Min,
Youngjeon Lee,
Jae-Won Huh,
Sang-Rae Lee,
Sang-Hyun Kim,
Deog-Bon Koo, Young-Kug Choo,
Hwan Mook Kim,
Sun-Uk Kim,
Kyu-Tae Chang
[show abstract]
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ABSTRACT: The coupling of autophagy and endoplasmic reticulum (ER) stress has been implicated in a variety of biological processes; however, little is known regarding the involvement of the autophagy/ER stress pathway in early embryogenesis or the underlying mechanism(s). Here, we showed that the developmental competence of in vitro-produced (IVP) bovine embryos was highly dependent on the autophagy/ER stress balance. Although relative abundances of autophagy-associated gene transcripts, including LC3, Atg5, and Atg7 transcripts, were high in oocytes and throughout the early stages of preattachment development, extensive autophagosome formation was only detected in fertilized embryos. Using an inducer and inhibitor of autophagy, we showed that transient elevation of autophagic activity during early preattachment development greatly increased the blastocyst development rate, trophectoderm cell numbers, and blastomere survival; these same parameters were reduced by both inhibition and prolonged induction of autophagy. Interestingly, the induction of autophagy reduced ER stress and associated damage, while the developmental defects in autophagy-inhibited embryos were significantly alleviated by ER stress inhibitor treatment, indicating that autophagy is a negative regulator of ER stress in early embryos. Collectively, these results suggest that early embryogenesis of IVP bovine embryos depends on an appropriate balance between autophagy and ER stress. These findings may increase our understanding of important early developmental events by providing compelling evidence concerning the tight association between autophagy and ER stress, and may contribute to the development of strategies for the production of IVP bovine blastocysts with high developmental competence.
Biology of Reproduction 04/2012; 87(1):8, 1-11. · 4.01 Impact Factor
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ABSTRACT: Gangliosides are ubiquitous membrane components in mammalian cells and are suggested to play important roles in various functions
such as cell-cell interaction, adhesion, cell differentiation, growth control and signaling. Among all ganglio-series gangliosides,
GM3 has the simplest carbohydrate structure, and has been shown as a major ganglioside in male reproductive system. To study
GM3 distribution in the seminiferous tubule and epididymis, frozen sections were stained with specific monoclonal antibody
(MAb) against ganglioside GM3. In the seminiferous tubule of testis, pachytene spermatocytes and spermatids expressed ganglioside
GM3, but not in spermatogonia and sertoli cells. Spermatogonia and sertoli cells near the basement membrane were negatively
reacted to anti-CM3. In the epididymis, GM3 was expressed only in some interstitial cells. Taken together, these results suggest
that the expression of ganglioside GM3 in rat seminiferous tubule and epididymis is spatio-temporally regulated during spermatogenesis.
Archives of Pharmacal Research 04/2012; 24(4):360-366. · 1.59 Impact Factor
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Zhe Lu,
Kyung-Jin Lee,
Yingxue Shao,
Jeong-Hwan Lee,
Yangkang So, Young-Kug Choo,
Doo-Byoung Oh,
Kyung-A Hwang,
Seung Han Oh,
Yeon Soo Han,
Kisung Ko
[show abstract]
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ABSTRACT: The tumor-associated antigen GA733 is a cell-surface glycoprotein highly expressed in colorectal carcinomas. In this study, 3 recombinant genes were constructed as follows: GA733 tagged to the ER retention sequence KDEL (GA733K), GA733 fused to the immunoglobulin Fc fragment (GA733-Fc), and GA733-Fc fused to the ER retention sequence (GA733-FcK). Agrobacterium-mediated transformation was used to generate transgenic plants expressing recombinant genes. The presence of transgenes was confirmed by genomic PCR. Western blot, confocal immunofluorescence, and sandwich ELISA showed the expression of recombinant proteins. The stability, flexibility, and bioactivity of recombinant proteins were analyzed and demonstrated through N-glycosylation analysis, animal trials, and sera ELISA. Our results suggest that the KDEL retained proteins in ER with oligomannose glycan structure and enhanced protein accumulation level. The sera of mice immunized with GA733-FcK purified from plants contained immunoglobulins which were at least as efficient as the mammalian-derived GA733-Fc at recognizing human colorectal cancer cell lines. Thus, a plant system can be used to express the KDEL fusion protein with oligomannose glycosylation, and this protein induces an immune response which is comparable to non-KDEL-tagged, mammalian-derived proteins.
Journal of Biomedicine and Biotechnology 01/2012; 2012:364240. · 2.44 Impact Factor
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ABSTRACT: The human colorectal carcinoma-associated GA733 antigen epithelial cell adhesion molecule (EpCAM) was initially described as a cell surface protein selectively expressed in some myeloid cancers. Gangliosides are sialic acid-containing glycosphingolipids involved in inflammation and oncogenesis. We have demonstrated that treatment with anti-EpCAM mAb and RAW264.7 cells significant inhibited the cell growth in SW620 cancer cells, but neither anti-EpCAM mAb nor RAW264.7 cells alone induced cytotoxicity. The relationship between ganglioside expression and the anti- cancer effects of anti-EpCAM mAb and RAW264.7 was investigated by high-performance thin-layer chromatography. The results demonstrated that expression of GM1 and GD1a significantly increased in the ability of anti-EpCAM to inhibit cell growth in SW620 cells. Anti-EpCAM mAb treatment increased the expression of anti-apoptotic proteins such as Bcl-2, but the expression of pro-apoptotic proteins Bax, TNF-α, caspase-3, cleaved caspase-3, and cleaved caspase-8 were unaltered. We observed that anti-EpCAM mAb significantly inhibited the growth of colon tumors, as determined by a decrease in tumor volume and weight. The expression of anti-apoptotic protein was inhibited by treatment with anti-EpCAM mAb, whereas the expression of pro-apoptotic proteins was increased. These results suggest that GD1a and GM1 were closely related to anticancer effects of anti-EpCAM mAb. In light of these results, further clinical investigation should be conducted on anti-EpCAM mAb to determine its possible chemopreventive and/or therapeutic efficacy against human colon cancer.
Experimental and Molecular Medicine 12/2011; 43(12):693-701. · 2.48 Impact Factor
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Dong Hoon Kwak,
Jung-Woo Jin,
Jae-Sung Ryu,
Kinram Ko,
So-Dam Lee,
Jeong-Woong Lee,
Ji-Su Kim,
Kyu Yong Jung,
Kisung Ko,
Jin Yeul Ma,
Kyung-A Hwang,
Kyu-Tae Chang, Young-Kug Choo
[show abstract]
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ABSTRACT: Gangliosides play an important role in neuronal differentiation processes. The regulation of ganglioside levels is related to the induction of neuronal cell differentiation. In this study, the ST8Sia5 gene was transfected into mESCs and then differentiated into neuronal cells. Interestingly, ST8Sia5 gene transfected mESCs expressed GQ1b by HPTLC and immunofluorescence analysis. To investigate the effects of GQ1b over-expression in neurogenesis, neuronal cells were differentiated from GQ1b expressing mESCs in the presence of retinoic acid. In GQ1b expressing mESCs, increased EBs formation was observed. After 4 days, EBs were co-localized with GQ1b and nestin, and GFAP. Moreover, GQ1b co-localized with MAP-2 expressing cells in GQ1b expressing mESCs in 7-day-old EBs. Furthermore, GQ1b expressing mESCs increased the ERK1/2 MAP kinase pathway. These results suggest that the ST8Sia5 gene increases ganglioside GQ1b and improves neuronal differentiation via the ERK1/2 MAP kinase pathway.
BMB reports 12/2011; 44(12):799-804. · 1.72 Impact Factor
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ABSTRACT: MicroRNAs (miRNAs) are small non-coding RNAs that regulate diverse biological processes. We cloned novel small RNA from human mesenchymal stem cells (hMSCs) and termed microRNA-5787 (hsa-miR-5787) that met the criteria for a miRNA. The level of miR-5787 was elevated in senescent fibroblasts. Based on the target prediction algorithm and results that were obtained, we find that eukaryotic translation initiation factor 5 (eIF5) is a target of miR-5787. Similar to the over-expression of miR-5787, we showed that repression of eIF5 in fibroblasts negatively affected cell growth. Therefore, we propose that the miR-5787 represses cell growth, in part, by targeting eIF5.
Biochemical and Biophysical Research Communications 10/2011; 415(4):567-72. · 2.48 Impact Factor
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ABSTRACT: The aim of the present study was to investigate the effects of oral administration of the insulin-like growth factor-I-rich fraction (IGF-I-RF) from bovine colostral whey on the regulation of blood glucose levels in streptozotocin (STZ)-induced diabetic mice. We obtained a peptide fraction containing IGF-I (10 ng/mg protein) from Holstein colostrum within 24 h after parturition by using ultrafiltration. The blood glucose levels of STZ-induced diabetic mice fed with IGF-I-RF (50 μg/kg per d) were significantly reduced by 11 and 33 % at weeks 2 and 4, respectively (P < 0·05). The body weights of STZ-induced diabetic mice increased following the oral administration of the IGF-I-RF. The kidney weights of STZ-induced diabetic mice decreased significantly (P < 0·05) following the administration of the IGF-I-RF, and the liver weights of STZ-induced diabetic mice decreased significantly (P < 0·05) following the administration of 50 μg/kg per d of the IGF-I-RF. The present results indicate that the IGF-I-RF obtained from Holstein colostrum could be a useful component for an alternative therapeutic modality for the treatment of diabetes in insulin-resistant patients.
The British journal of nutrition 10/2011; 108(1):39-45. · 3.45 Impact Factor
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ABSTRACT: In this study we investigated for the first time the transcriptional regulation of pig Galβ1,3GalNAc α2,3-sialyltransferase (pST3Gal I) in response to TGF-β1 in porcine kidney PK-15 cells. The pST3Gal I gene was found to span about 90kb and to be composed of 8 exons including 2 exons in the 5'-untranslated region. RT-PCR analysis indicated that the induction of pST3Gal I by TGF-β1 is regulated at the transcriptional level. Functional analysis of the 5'-flanking region of the pST3Gal I gene revealed the -1257 to -976 region functions as the TGF-β1-inducible promoter and that the Smad-binding site at -1020 is crucial for TGF-β1-induced expression of pST3Gal I in PK-15 cells. In addition, the transcriptional activity of pST3Gal I induced by TGF-β1 in PK-15 cells was strongly inhibited by SIS3, which is a specific Smad-3 inhibitor. In summary, our results identified the core promoter region in the pST3Gal I promoter and demonstrated that Smad-3 binding to the Samd-3 binding site at -1020 is essential for transcriptional activation of pST3Gal I in TGF-β1-induced PK-15 cells.
Biochemical and Biophysical Research Communications 09/2011; 414(1):159-64. · 2.48 Impact Factor
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Bong-Seok Song,
Ji-Su Kim,
Seung-Bin Yoon,
Kyu-Sun Lee,
Deog-Bon Koo,
Dong-Seok Lee, Young-Kug Choo,
Jae-Won Huh,
Sang-Rae Lee,
Sun-Uk Kim,
Sang-Hyun Kim,
Hwan Mook Kim,
Kyu-Tae Chang
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ABSTRACT: Somatic cell nuclear transfer (SCNT) is a powerful tool, not only for producing cloned animals, but also in revealing various early developmental events. However, relatively little is known regarding the biological events and underlying mechanism(s) directly associated with early development of SCNT embryos. Here, we show that production of high-quality bovine SCNT blastocysts is dependent on the method used for fusion and the associated reduction in endoplasmic reticulum (ER) stress. During fusion between the donor cell and the enucleated oocyte, electrofusion triggers spontaneous oocyte activation, accompanied by an increase in intracellular Ca(2+) and improper nuclear remodelling. These events can be greatly reduced by the use of Sendai virus (SV)-mediated fusion. Moreover, SV-SCNT improves the blastulation rate and blastocyst quality, defined by the number and ratio of inner cell mass and trophectoderm cells in each blastocyst, in comparison with electrofusion-mediated SCNT (E-SCNT). Interestingly, expression of ER-stress-associated genes and blastomere apoptosis were significantly increased in E-SCNT embryos. These increases could be reversed by inhibition of ER stress or by using the SV-mediated fusion method. Collectively, these results indicate that SV-mediated fusion improves the developmental competence and quality of SCNT blastocysts, by reducing ER-stress-associated apoptosis.
Reproduction Fertility and Development 08/2011; 23(6):826-36. · 2.11 Impact Factor
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ABSTRACT: Gangliosides have been suggested to play important roles in various functions such as adhesion, cell differentiation, growth control, and signaling. Mouse follicular development, ovulation, and luteinization during the estrous cycle are regulated by several hormones and cell-cell interactions. In addition, spermatogenesis in seminiferous tubules of adult testes is also regulated by several hormones, including follicle-stimulating hormone (FSH) and luteinizing hormone (LH) and cell-cell interactions. The regulation of these processes by hormones and cell-cell interactions provides evidence for the importance of surface membrane components, including gangliosides. During preimplantation embryo development, a mammalian embryo undergoes a series of cleavage divisions whereby a zygote is converted into a blastocyst that is sufficiently competent to be implanted in the ma ternal uterus and continue its development. Mouse embryonic stem (mES) cells are pluripotent cells derived from mouse embryo, specifically, from the inner cell mass of blastocysts. Differentiated neuronal cells are derived from mES cells through the formation of embryonic bodies (EBs). EBs recapitulate many aspects of lineage-specific differentiation and temporal and spatial gene expression patterns during early embryogenesis. Previous studies on ganglioside expression during mouse embryonic development (including during in vitro fertilization, ovulation, spermatogenesis, and embryogenesis) reported that gangliosides were expressed in both undifferentiated and differentiated (or differentiating) mES cells. In this review, we summarize some of the advances in our understanding of the functional roles of gangliosides during the stages of mouse embryonic development, including ovulation, spermatogenesis, and embryogenesis, focusing on undifferentiated and differentiated mES cells (neuronal cells).
Experimental and Molecular Medicine 06/2011; 43(7):379-88. · 2.48 Impact Factor
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Ji-Su Kim,
Bong-Seok Song,
Sang-Rae Lee,
Seung-Bin Yoon,
Jae-Won Huh,
Sun-Uk Kim,
Ekyun Kim,
Sang-Hyun Kim, Young-Kug Choo,
Deog-Bon Koo,
Kyu-Tae Chang
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ABSTRACT: Metaphase II oocyte production was significantly increased by treatment with E(2) during the first half of the total in vitro maturation (IVM) period, which was further evidenced by an increase in monospermic fertilization, blastocyst formation, or blastomere viability of IVF- or somatic cell nuclear transfer-derived embryos. Thus, we concluded that transient E(2) supplementation could improve the IVM rate and subsequent developmental competence in pigs.
Fertility and sterility 04/2011; 95(8):2582-4. · 3.97 Impact Factor
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ABSTRACT: In this study, we investigated the regulatory role of ganglioside GD1a in the differentiation of osteoblasts from human mesenchymal stem cells (hMSCs) by using lentivirus-containing short hairpin (sh)RNA to knockdown ST3 β-galactoside α-2, 3-sialyltransferase 2 (ST3Gal II) mRNA expression. After hMSCs were infected for 72 h with the lentivirus constructed with ST3Gal II shRNAs, the puromycin-resistant cells were selected and subcultured to produce hMSCs with ST3Gal II mRNA knockdown. The hMSCs established from human dental papilla abundantly expressed CD44 and CD105, but not CD45 and CD117. Osteoblasts that differentiated from normal hMSCs showed a significant increase in alkaline phosphatase (ALP) activity and ganglioside GD1a expression level compared with those in hMSCs. Lentiviral infection of hMSCs successfully induced a marked inhibition of ST3Gal II mRNA expression and caused a significant decrease in ALP activity and ganglioside GD1a expression. During osteoblastic differentiation, the increased ALP activity remarkably reduced by suppression of ganglioside GD1a expression by ST3Gal II shRNA. Ganglioside GD1a and ALP were mainly expressed in the cell body of hMSCs and osteoblasts with colocalization. The phosphorylation of extracellular signal-regulated kinases (ERK) 1/2 mitogen-activated protein (MAP) kinase and epidermal growth factor receptor (EGFR) was significantly reduced in the osteoblasts that had differentiated from the hMSCs with ST3Gal II mRNA knockdown. These results suggest that ganglioside GD1a plays an important role in the regulation of osteoblastic differentiation of hMSCs through the activation of ERK 1/2 MAP kinase and EGFR.
Embryologia 04/2011; 53(3):323-32. · 2.21 Impact Factor
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Mira Song,
Da-Young Park,
Youngkwan Kim,
Kyung-Jin Lee,
Zhe Lu,
Kinarm Ko, Young Kug Choo,
Yeon Soo Han,
Mi-Hyun Ahn,
Doo-Byoung Oh,
Kisung Ko
[show abstract]
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ABSTRACT: Advantages of the baculovirus insect cell expression system for production of recombinant proteins include high capacity, flexibility, and glycosylation capability. In this study, this expression system was exploited to produce anti-cancer monoclonal antibody (mAb) CO17-1A, which recognizes the antigen GA733. The heavy chain (HC) and light chain (LC) genes of mAb CO17-1A were cloned under the control of P(10) and Polyhedrin promoters in the pFastBac dual vector, respectively. Gene expression cassettes carrying the HC and LC genes were transposed into a bacmid in Escherichia coli (DH10Bac). The transposed bacmid was transfected to Sf9 insect cells to generate baculovirus expressing mAb CO17-1A. Confocal immunofluorescence and Western blot analyses confirmed expression of mAb CO17-1A in baculovirus-infected insect cells. The optimum conditions for mAb expression were evaluated at 24, 48, and 72 h after the virus infection at an optimum virus multiplicity of infection of 1. Expression of mAb CO17-1A in insect cells significantly increased at 72 h after infection. HPLC analysis of glycosylation status revealed that the insect-derived mAb (mAb(I)) CO17-1A had insect specific glycan structures. ELISA showed that the purified mAb(I) from cell culture supernatant specifically bound to SW948 human colorectal cancer cells. Fluorescence-activated cell sorting analysis showed that, although mAb(I) had insect specific glycan structures that differed from their mammalian counterparts, mAb(I) similarly interacted with CD64 (FcgammaRI) and Fc of IgG, compared to the interactions of mammalian-derived mAb. These results suggest that the baculovirus insect cell expression system is able to express, assemble, and secrete biofunctional full size mAb.
Journal of Bioscience and Bioengineering 08/2010; 110(2):135-40. · 1.79 Impact Factor
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Animal Cells and Systems. 06/2010; 14(2):83-89.
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ABSTRACT: Human adipose-derived stem cells (hADSCs) and dental pulp-derived stem cells (hDPSCs) have been considered alternative sources of adult stem cells because of their potential to trans-differentiate into multiple cell lineages. This study investigated the possible role of gangliosides in the osteoblast differentiation of hADSCs and hDPSCs. First, we investigated characterization of hADSCs and hDPSCs using FACS analysis. Mesenchymal stem cell specific markers, CD44 and CD105, were expressed but not hematopoietic markers, CD45 and CD117 in both of hADSCs and hDPSCs. High-performance thin-layer chromatography analysis showed that increased gangliosides were associated with differentiation of hADSCs and hDPSCs into osteoblasts. RT-PCR analysis confirmed that osteoblast specific genes, ALP, BMP-2, collagen were expressed in differentiated osteoblasts, however, the another osteoblast specific gene, osteocalcin, was not expressed. When hADSCs and hDPSCs were cultured under osteoblast-differentiation conditions, alkaline phosphatase (ALP) activity was increased in comparison to hADSCs and hDPSCs. Furthermore, specifically both ALP activity and ganglioside expression increased more in hDPSCs-derived osteoblasts than hADSCs-derived osteoblasts. These results suggest that gangliosides play a more important role in regulating the osteoblast-differentiation of hDPSCs compared to hADSCs.
Archives of Pharmacal Research 04/2010; 33(4):585-91. · 1.59 Impact Factor
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ABSTRACT: This study examined the estrogenic activity produced by aqueous extracts of silkworm (Bombyx mori) pupae in ovariectomized (OVX) rats. The components of silkworm pupae were extracted in distilled water at room temperature for 6 hours. The ovaries of six-week old female rats were then bilaterally removed. One week after OVX, the animals were treated with 200, 400 or 600 mg/kg/day of silkworm pupae extracts. The body weights of the OVX rats increased remarkably compared to the control rats, however their relative uterus weights to body weights decreased significantly. Treatment with the aqueous extracts of silkworm pupae dramatically improved the decreased uterus weights of OVX rats, with the highest increase observed in treatment with 200 mg/kg/day of the aqueous extracts. Additionally, treatment with aqueous extracts (200 mg/kg/day) of silkworm pupae significantly elevated the serum 17beta-estradiol contents of OVX rats when compared to the control animals. To examine the toxic effects of silkworm pupae on the hepatic functions of OVX rats, the levels of serum glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT) were measured. The serum GOT and GPT levels did not change in response to the administration of aqueous extracts (200, 400 and 600 mg/kg/day) for 4-weeks. Taken together, these results suggest that the aqueous extracts of silkworm pupae may have estrogenic activity, which suggests that silkworm pupae may be useful in the prevention and/or treatment of menopausal disorders caused by deficiencies in female sexual hormones, including estrogen.
The American Journal of Chinese Medicine 01/2010; 38(1):89-97. · 1.98 Impact Factor
