• Journal of Physiology & Pathology in Korean Medicine
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• v.23 no.6
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• pp.1299-1304
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• 2009

We have previously shown that water extract of deer antler (WEDA) inhibited RANKL-mediated osteoclast differentiation from bone marrow macrophages by suppressing c-Fos and NFATc1 expression. Thus, we examined the effect of WEDA in inflammation-induced bone loss in vivo. Here we found that WEDA inhibited osteoblast-supported osteoclast differentiation induced by lipopolysaccharide (LPS). However, WEDA did not suppress the expression of receptor activator of NF-${\kappa}B$ ligand (RANKL) in response to LPS in osteoblasts. WEDA also inhibited the bone resorptive activity of mature osteoclasts. To examine the effect of WEDA on bone loss, when LPS injected subcutaneously in mice, bone loss was greatly increased, but WEDA treatment inhibited LPS-mediated bone loss. Taken together, we conclude that WEDA inhibited osteoclast differentiation and bone resorption in vitro and in vivo. Thus WEDA may be useful in the treatment of bone-related disorders.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.25-34
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• 2003

Human thrombopoietin (hTPO) is a cytokine that plays a central role in megakaryopoiesis. To direct hTPO expression in the mammary gland, an expression vector was constructed by combining the promoter of bovine beta-casein gene, cDNA of hTPO and neomycin resistance gene (pBT-L neo). Fibroblast cells derived from cow's ear skin tissue were transfected with the expression vector (pBT-L neo) using Lipofectamine. Transfected cells resistant to G418 trea？nt were cultured to form the colonies for more than 2 weeks. The transformed colonies identified by PCR were further expanded prior to nuclear transfer. Reconstructed oocytes with transformed cells were electrofused, activated using calcium ionophore and 6-DMAP, and cultured in vitro for 7 days. Of 35 cell colonies analyzed by PCR, 29 colonies (82.9%) were positive for the hTPO gene. Cleavage and developmental rates to the blastocyst stage of reconstructed embryos with the transformed cells were 65.1% and 23.8%, respectively Of 29 blastocysts that developed from reconstructed embryos with the transformed cells, 27 embryos (93.1%) were transgenic. These results indicate that transgenic bovine embryos can be efficiently produced by somatic cell nuclear transfer using transformed cells.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.4
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• pp.501-510
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• 2007

The present study was conducted to establish primary bovine muscle satellite cell (MSC) culture conditions and to investigate the effects of various steroid hormones on transcription of the genes involved in muscle cell proliferation and differentiation. Of three different types of proteases (type II collagenase, pronase and trypsin-EDTA) used to hydrolyze the myogenic satellite cells from muscle tissues, trypsin-EDTA treatment yielded the highest number of cells. The cells separated by hydrolysis with type II collagenase and incubated on gelatin-coated plates showed an enhanced cell attachment onto the culture plate and cell proliferation at an initial stage of cell growth. In this study, the bovine MSCs were maintained in vitro up to passage 16 without revealing any significant morphological change, and even to when the cells died at passage 21 with decreased or almost no cell growth or deformities. When the cells were incubated in a steroid-depleted environment (DMEM(-)/10% CDFBS (charcoal-dextran stripped FBS)), they grew slowly initially, and were widened and deformed. In addition, when the cells were transferred to an incubation medium containing steroid (DMEM(+)/10% FBS), the deformed cells resumed their growth and returned to a normal morphology, suggesting that steroid hormones are crucial in maintaining normal MSC morphology and growth. The results demonstrated that treatments with 19-nortestosterone and testosterone significantly increased AR gene expression (p<0.05), implying that both testosterone and 19-nortestosterone bind with AR and that the hormone bound-AR complex up-regulates the genes of its own receptor (AR) plus other genes involved in satellite cell growth and differentiation in bovine muscle.

• Journal of Korean Medicine Rehabilitation
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• v.30 no.4
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• pp.1-16
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• 2020

Objectives The main purpose of this study was to evaluate the bone healing effect of Pahyeolsandong-tang (PHT)(Poxiesanteng-tang) extract in tibia fracture-induced mice. Methods PHT was extracted using a solution of 35% ethanol in 60℃ for 8 hours. Mice were randomly divided into 4 groups (normal, control, PHT 50 and PHT 100). Mice of experimental groups were medicated with PHT 50 or 100 mg/kg for 7 to 21 days. To clarify the effect of bone fracture healing, relative messenger RNA (mRNA) expressions of osteocalcin (OCN), runt-related transcription factor 2 (Runx2), osterix (OSX), Sox9, collagen type II alpha 1 chain (Col2a1), receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG) were examined. Results In in vitro experiment, relative mRNA expression of OCN, Runx2, Col2a1 was significantly increased in PHT treated group to compare with control differentiation group. In in vivo experiment, relative mRNA expression of OCN, Runx2, OSX, Sox9, Col2a1, RANKL, OPG was significantly increased in PHT treated group. Conclusions This study showed that PHT accelerates bone fracture healing through the activation of osteoclasts and osteoblasts. It was showed that PHT significantly promotes osteoblasts differentiation by osteoblast differentiation markers such as OCN, Runx2, Col1a2. Also it was investigated that PHT had stimulatory effect on osteoblasts function through enhancing OCN, Runx2, OSX, Sox9, Col2a1 and, osteoclasts function through enhancing RANKL and OPG markers. PHT effectively promotes bone fracture healing process through activation of osteoblasts and osteoclasts.

• Journal of Plant Biotechnology
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• v.44 no.2
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• pp.164-170
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• 2017

A rapid and efficient in vitro regeneration system was established for Hibiscus syriacus L. The successful regeneration protocol employs induction of shoot organogenesis on leaf, petiole, and root explants. Among the various plant growth regulators evaluated, thidiazuron (TDZ) was the most effective for inducing rapid shoot formation. Most efficient shoot regeneration frequency was obtained from Murashige and Skoog (MS) media containing 0.01 mg/L TDZ. Regeneration efficiency was highest in the roots, and lowest in the leaves. A combination of 0.01 mg/L TDZ with benzyladenine (BAP) markedly improved the frequency of shoot differentiation from the root (up to 98%) and petiole (up to 88%) explants. Furthermore, leaf and petiole explants showed the highest frequency of shoot induction in half-strength MS media containing 0.01 mg/L TDZ and 1.0 mg/L BAP, while root explants formed the greatest number of shoots when 0.01 mg/L TDZ and 0.1 mg/L BAP were added to half-strength MS media. Although the frequency of shoot differentiation from leaf explants was only 50%, the leaf is considered the most efficient plant organ for use in tissue culture because leaves are easier to obtain than roots and petioles. Our findings show that various organs of H. syriacus can be used for plant regeneration, and the protocol developed in this study may be applicable in the horticulture industry.

• Journal of Physiology & Pathology in Korean Medicine
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• v.25 no.1
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• pp.109-114
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• 2011

Atractylodis Rhizoma Alba is commonly used herbal medicine and it has been known that has immuno-regualtory effects and anti-cancer effects. The inhibition of osteoclastogenesis is essential for the prevention and treatment of osteoporosis. The aim of this study was to evaluate the effects of Atractylodis Rhizoma Alba on osteoclast differentiation in vitro and on resorbing activity of osteoclast. Osteoclast formation was evaluated in bone marrow cells (BMC) in the presence or absence of Atractylodis Rhizoma Alba. The expression of c-fos, tartrate-resistant acid phosphatase (TRAP), OSCAR, DC-STAMP, cathepsin K, MafB and NFATc1 mRNA in osteoclast precursor were assessed by RT-PCR. The levels of TNF receptor-associated factor-6 (TRAF-6), c-fos and NFATc1 protein were assessed by Western blot analysis. Also the correlation with MAPKs and NF-${\kappa}B$ pathways were measured by using Western blot analysis. With bone resorption study, I tried to evaluate the inhibitory effects of Atractylodis Rhizoma Alba on mature osteoclast function. Atractylodis Rhizoma Alba inhibited the RANKL induced osteoclastic differentiation from bone marrow macrophage in a dose dependant manner without cellular toxicity. Gene expression of c-fos and NFATc1 was significantly down regulated with Atractylodis Rhizoma Alba treatment. Atractylodis Rhizoma Alba markedly inhibited the RANKL-induced osteoclastogenesis through suppression of nuclear factor kappa b (NF-${\kappa}B$) pathway, down stream pathway of p38, ERK and JNK pathway. Taken together, I concluded that Atractylodis Rhizoma Alba have beneficial effect on osteoporosis by inhibition of osteoclast differentiation and by inhibition of functioning osteoclast. Thus I expect that Atractylodis Rhizoma Alba could be a treatment option for osteoporosis.

• BMB Reports
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• v.54 no.10
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• pp.505-515
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• 2021

Human pluripotent stem cells (hPSCs) include human embryonic stem cells (hESCs) derived from blastocysts and human induced pluripotent stem cells (hiPSCs) generated from somatic cell reprogramming. Due to their self-renewal ability and pluripotent differentiation potential, hPSCs serve as an excellent experimental platform for human development, disease modeling, drug screening, and cell therapy. Traditionally, hPSCs were considered to form a homogenous population. However, recent advances in single cell technologies revealed a high degree of variability between individual cells within a hPSC population. Different types of heterogeneity can arise by genetic and epigenetic abnormalities associated with long-term in vitro culture and somatic cell reprogramming. These variations initially appear in a rare population of cells. However, some cancer-related variations can confer growth advantages to the affected cells and alter cellular phenotypes, which raises significant concerns in hPSC applications. In contrast, other types of heterogeneity are related to intrinsic features of hPSCs such as asynchronous cell cycle and spatial asymmetry in cell adhesion. A growing body of evidence suggests that hPSCs exploit the intrinsic heterogeneity to produce multiple lineages during differentiation. This idea offers a new concept of pluripotency with single cell heterogeneity as an integral element. Collectively, single cell heterogeneity is Janus-faced in hPSC function and application. Harmful heterogeneity has to be minimized by improving culture conditions and screening methods. However, other heterogeneity that is integral for pluripotency can be utilized to control hPSC proliferation and differentiation.

• International Journal of Stem Cells
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• v.16 no.4
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• pp.425-437
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• 2023

Obesity, which continues to increase worldwide, was shown to irreversibly impair the differentiation potential and angiogenic properties of adipose tissue mesenchymal stromal cells (ADSCs). Because these cells are intended for regenerative medicine, especially for the treatment of inflammatory conditions, and the effects of obesity on the immunomodulatory properties of ADSCs are not yet clear, here we investigated how ADSCs isolated from former obese subjects (Ex-Ob) would influence macrophage differentiation and polarization, since these cells are the main instructors of inflammatory responses. Analysis of the subcutaneous adipose tissue (SAT) of overweight (OW) and Ex-Ob subjects showed the maintenance of approximately twice as many macrophages in Ex-Ob SAT, contained within the CD68⁺/FXIII-A^- inflammatory pool. Despite it, in vitro, coculture experiments revealed that Ex-Ob ADSCs instructed monocyte differentiation into a M2-like profile, and under inflammatory conditions induced by LPS treatment, inhibited HLA-DR upregulation by resting M0 macrophages, originated a similar percentage of TNF-α⁺ cells, and inhibited IL-10 secretion, similar to OW-ADSCs and BMSCs, which were used for comparison, as these are the main alternative cell types available for therapeutic purposes. Our results showed that Ex-Ob ADSCs mirrored OW-ADSCs in macrophage education, favoring the M2 immunophenotype and a mixed (M1/M2) secretory response. These results have translational potential, since they provide evidence that ADSCs from both Ex-Ob and OW subjects can be used in regenerative medicine in eligible therapies. Further in vivo studies will be fundamental to validate these observations.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.9
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• pp.1288-1302
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• 2011

The aim of this study was to analyze the proteome expression of bovine satellite cells from longissimus dorsi (LD), deep pectoral (DP) and semitendinosus (ST) muscle depots during in vitro myogenic differentiation. Proteomic profiling by twodimensional gel electrophoresis and mass spectrometry of differentiating satellite cells revealed a total of 38 proteins that were differentially regulated among the three depots. Among differentially regulated proteins, metabolic proteins like lactate dehydrogenase (LDH), malate dehydrogenase (MDH) were found to be up regulated in ST, while alpha-enolase (NNE) in LD and DP depot satellite cells were down regulated. Also, our analysis found that there was a prominent up regulation of cytoskeletal proteins like actin, actincapping protein and transgelin along with chaperone proteins like heat shock protein beta 1 (HSPB 1) and T-complex protein 1 (TCP-1). Among other up regulated proteins, LIM domain containing protein, annexin 2 and Rho GDP-dissociation inhibitor 1 (Rho GDI) are observed, which were already proven to be involved in the myogeneis. More interestingly, satellite cells from ST depot were found to have a higher myotube formation rate than the cells from the other two depots. Taken together, our results demonstrated that, proteins involved in glucose metabolism, cytoskeletal modeling and protein folding plays a key role in the myogenic differentiation of bovine satellite cells.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.30 no.5
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• pp.438-443
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• 2004

Objectives : The proper development of the facial structures relies upon a sequence of tightly regulated signaling interactions between the ectoderm and mesoderm involving the participation of several families of signaling molecules. Among these, bone morphogenetic proteins (BMPs) have been suggested to be a key signal that regulates the development of the mandible and the initiation and morphogenesis of the teeth. The aim of this study was to examine the artificial development of the mandibular structures and to examine the role of BMPs on tooth morphogenesis and differentiation using an organ culture system. Materials and Methods : The tooth germs from Ed 11.5, 13.5 mice were dissected, and transplanted into the diastema of the mandible primordia. The mandibles containing the transplanted tooth germs were cultured in vitro. During this period, beads soaked with BMP4 were implanted around the transplanted tooth germs. In addition, a diastema block containing the transplanted tooth germ was dissected, then transferred to an adult mouse kidney. After the organ culture, the developing mandibular explant was removed from the kidney and prepared for the tissue specimens. Odontogeneis of the transplanted tooth germs was examined after Hematoxylin-eosin, Masson-trichrome staining. Results : Proliferation and differentiation of the tooth germs cultured in the diastema was observed. In the BMP4-treated tooth germs, the formation of the first and second molars was noted. The crown of the developing tooth showed the formation of a mature cusp with the deposition of enamel and dentin matrix. In conclusion, it was confirmed that BMP4 is involved in the formation of a dental crown and the differentiation of ameloblasts and odontoblasts of the molar tooth during the development of the transplanted tooth germs.