• Development and Reproduction
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• v.12 no.3
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• pp.221-229
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• 2008

For the clinical application, it is needed to keep characteristics of stem cells after storage for a long time. In the present study, we examined stem cell properties of human cord-derived stem cells (HUC) after cryopreservation. Cells were isolated from human umbilical cord and cultured in vitro. At passage 2 or 3, HUC were suspended at a concentration of $1.0{\times}10^6/m{\ell}$ in cryomedium consisting of DMSO and FBS. After freezing at $-80^{\circ}C$ overnight, HUC were cryopreserved at $-196^{\circ}C$ nitrogen gas. After 6 months, HUC were thawed and cultured in vitro. Assessment for the stem cell properties was made upon the morphology, population doubling time, and expression profiles of genes and various proteins. Cryopreserved HUC showed more than 70% viability and maintained fibroblast-like morphology similar to HUC before cryopreservation. Throughout the culture, they underwent average 42.8 doublings and produced $6.75{\times}{10^{18}}$ cells. RT-PCR analyses showed that cryopreserved HUC expressed Oct-4, nanog, SCF, NCAM, nestin, GATA-4, BMP4, and HLA-1 genes. They did not express Brachyury and HLA-DR genes. Immunocytochemical studies showed that cryopreserved HUC reacted with antibodies against SSEA-3, -4, Thy-1, vimentin, fibronectin, HCAM, ICAM, HLA-1 proteins. They did not react with antibody against HLA-DR protein. Theses genes and proteins expression patterns of cryopresserved HUC were similar to those of HUC before cryopreservation. These results suggest that cryopreserved HUC could retain proliferative potential and they expressed various genes and proteins similar to HUC before cryopreservation. Thus, cryopreservation might be useful for HUC for future research and clinical application.

• Journal of Animal Science and Technology
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• v.65 no.1
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• pp.16-31
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• 2023

Cultured meat is a potential sustainable food generated by the in vitro myogenesis of muscle satellite (stem) cells (MSCs). The self-renewal and differentiation properties of MSCs are of primary interest for cultured meat production. MSC proliferation and differentiation are influenced by a variety of growth factors such as insulin-like growth factors (IGF-1 and IGF-2), transforming growth factor beta (TGF-β), fibroblast growth factors (FGF-2 and FGF-21), platelet-derived growth factor (PDGF) and hepatocyte growth factor (HGF) and by hormones like insulin, testosterone, glucocorticoids, and thyroid hormones. In this review, we investigated the roles of growth factors and hormones during cultured meat production because these factors provide signals for MSC growth and structural stability. The aim of this article is to provide the important idea about different growth factors such as FGF (enhance the cell proliferation and differentiation), IGF-1 (increase the number of myoblasts), PDGF (myoblast proliferation), TGF-β1 (muscle repair) and hormones such as insulin (cell survival and growth), testosterone (muscle fiber size), dexamethasone (myoblast proliferation and differentiation), and thyroid hormones (amount and diameter of muscle fibers and determine the usual pattern of fiber distributions) as media components during myogenesis for cultured meat production.

• Development and Reproduction
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• v.12 no.1
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• pp.87-95
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• 2008

Neural stem/precursor derived from pluripotent human embryonic stem cells (hESCs) has considerable therapeutic potential due to their ability to generate various neural cells which can be used in cell-replacement therapies for neurodegenerative diseases. However, production of neural cells from hESCs remains technically very difficult. Understanding neural-tube like rosette characteristic neural precursor cells from hESCs may provide useful information to increase the efficiency of hESC neural differentiation. Generally, neural rosettes were derived from differentiating hEBs in attached culture system, however this is time-consuming and complicated. Here, we examined if neural rosettes could be formed in suspension culture system by bypassing attachment requirement. First, we tested whether the size of hESC clumps affected the formation of human embryonic bodies (hEBs) and neural differentiation. We confirmed that hEBs derived from $500{\times}500\;{\mu}m$ square sized hESC clumps were effectively differentiated into neural lineage than those of the other sizes. To induce the rosette formation, regular size hEBs were derived by incubation of hESC clumps($500{\times}500\;{\mu}m$) in EB medium for 1 wk in a suspended condition on low attachment culture dish and further incubated for additional $1{\sim}2$ wks in neuroectodermal sphere(NES)-culture medium. We observed the neural tube-like rosette structure from hEBs after $7{\sim}10$ days of differentiation. Their identity as a neural precursor cells was assessed by measuring their expressions of neural precursor markers(Vimentin, Nestin, MSI1, MSI2, Prominin-1, Pax6, Sox1, N-cadherin, Otx2, and Tuj1) by RT-PCR and immunofluorescence staining. We also confirmed that neural rosettes could be terminally differentiated into mature neural cell types by additional incubation for $2{\sim}6$ wks with NES medium without growth factors. Neuronal(Tuj1, MAP2, GABA) and glial($S100{\beta}$ and GFAP) markers were highly expressed after $2{\sim}3$ and 4 wks of incubation, respectively. Expression of oligodendrocyte markers O1 and CNPase was significantly increased after $5{\sim}6$ wks of incubation. Our results demonstrate that rosette forming neural precursor cells could be successfully derived from suspension culture system and that will not only help us understand the neural differentiation process of hESCs but also simplify the derivation process of neural precursors from hESCs.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.3
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• pp.186-196
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• 2010

Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.5
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• pp.587-594
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• 2000

At present embryonic stem (ES) cells with confirmed pluripotential properties are only available in the mouse. Recently, we were able to isolate, culture and genetically transform primordial germ cell (PGC)-derived cells from pig embryos and demonstrate their ability to contribute to chimera development in the pig. In order to determine whether the system we developed could be used to isolate embryonic germ (EG) cells from other mammalian species, we placed isolated PGCs from cattle, goats, rabbits and rats in culture. Briefly, PGCs were isolated from fetuses of cow (day 30-50), goat (day 25), rabbit (day 15-18) and rat (day 11-12), and plated on STO feeder cells in Dulbecco's modified Eagle's medium (DMEM): Ham's F10 medium (1:1) supplemented with 0.01 mM nonessential amino acids, 2 mM L-glutamine, 0.1 mM $\beta$ - mercaptoethnol, soluble recombinant human stem cell factor (SCF; 40ng/ml), human basic fibroblast growth factor (bFGF; 20ng/ml) and human leukemia inhibitory factor (LIF; 20ng/ml). For maintenance of the cells, colonies were passed to fresh feeders every 7-10 days. In all species tested, we were able to obtain and maintain colonies with ES-like morphology. Their developmental potential was tested by alkaline phosphatase (AP) staining and in vitro differentiation assay. For genetic transformation, cells were electroporated with a construct containing the green fluorescent protein (GFP) under the control of the cytomegalovirus (CMV) promoter. GFP-expressing colonies were detected in cattle, rabbits and rats. These results suggest that PGC-derived cells from cattle, goats, rabbits and rats can be isolated, cultured, and genetically transformed, and provide the basis for analyzing their developmental potential and their possible use for the precise genetic modification of these species.

• CELLMED
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• v.4 no.1
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• pp.2.1-2.17
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• 2014

The plant kingdom is considered to be a repository of modern medicine, attributable to their rich source of bio-active molecules and secondary metabolites. It is indeed the Nutraceuticals that enhance immunity and ensure a healthier life because of their prophylactic and therapeutic values. Over centuries, papaya [Caricaceae; (Carica papaya Linn.)] is a renowned nutritious and medicinal plant. Each part of the papaya like root, stem, leaf, flower, fruit, seed, rinds, and latex has its own nutraceutical properties. It serves as food, cooking aid, and Ethnomedicine to prevent and treat wide-range of diseases and disorders. It has also been traditionally used as appetite enhancer, meat tenderizer, purgative, medicinal acne, abortifacient and vermifuge. Over decades, a series of scientific attempts were made to authenticate the nutraceutical properties of papaya. These studies validated that the papaya has antiplasmodial, antitrichochramal, antitrichomonal, antidengue, and anti-cancer activities. They have also exhibited that papaya possesses antiseptic, antiparasitic, anti-inflammatory, antidiabetic, and contraceptive features, and it helps in the management of sickle-cell anaemia, HIV, heart diseases and digestional disorders too. Nevertheless, the responsible bio-active molecules and their mode of actions remain indistinct and imprecise, and this calls for further pharmacological and clinical research on them. Conclusively, papaya is one of the naturally gifted plants; though its nutraceutical properties as a food or as a quasi-drug are poorly understood or undervalued by people. Accordingly, this scrutiny, demand for instigation of public health awareness campaigns to promote papaya consumption, so that the society shall acquire optimal benefits of papaya and in turn prevent and alleviate various diseases and illness.

• Journal of Korean Society of Forest Science
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• v.16 no.1
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• pp.33-62
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• 1972

Pitch pine (Pinus rigida Miller) in Korea has become one of the major silvicultural species for many years since it was introduced from the United States of America in 1907. To attain the more rational wood utilization basical researches on wood properties are primarily needed, since large scale of timber production from Pitch Pine trees has now been accomplishing in the forested areast hroughout the country. Under the circumustances, this experiment was carried out to study the wood anatomical, physical and mechanical properties of Pitch Pine grown in the country. Materials used in this study had been prepared by cutting the selected pitch pine trees from the Seoul National University Forests located in Suwon. To obtain and compare the anatomical and physical properties of the different parts of tree such as stem, branch, top and rootwood, this study had been divided into two categories (anatomical and physical). For the anatomical study macroscopical and microscopical features such as annual ring, intercellular cannal, ray, tracheid, ray trachid, ray parenchyma cell and pit etc. were observed and measured by the different parts (stem, branch, root and topwood) of tree. For the physical and mechanical properties the moisture content of geen wood, wood specific gravity, shrinkage, compression parallel to the grain, tension parallel and perpendicular to the grain, radial and tangential shear, bending, cleavage and hardness wree tested. According to the results this study may be concluded as follows: 1. The most important comparable features in general properties of wood among the different parts of tree were distinctness and width of annual ring, transition from spring to summerwood, wood color, odor and grain etc. In microscopical features the sizes of structural elements of wood were comparable features among the parts of tree. Among their features, length, width and thickness of tracheids, resin ducts and ray structures were most important. 2. In microscopical features among the different parts of tree stem and topwood were shown simillar reults in tissues. However in rootwood compared with other parts on the tangential surface distinctly larger ray structures were observed and measured. The maximum size of unseriate ray was attained to 27 cell ($550{\mu}$) height in length and 35 microns in width. Fusiform rays were formed occasionally the connected ray which contain one or several horizontal cannals. Branchwood was shown the same features like stemwood but the measured values were very low in comparing with other parts of tree. 3. Trachid length measured among the different parts of tree were shown largest in stem and shortest in branchwood. In comparing the tracheid length among the parts the differences were not shown only between stem and rootwood, but shown between all other parts of tree. Trachid diameters were shown widest in rootwood and narrowest in branchwood, and the differences among the different parts were not realized. Wall thickness were shown largest value in rootwood and smallest in branchwood, and the differences were shown between root and top or branchwood, and between stem and branch or top wood, but not shown between other parts of tree. 4. Moisture contents of green wood were shown highest in topwood and lowest in heartwood of stem. The differences among the different parts were recognized between top or heartwood and other parts of tree, but not between root and branchwood or root and sapwood. 5. Wood specific gravities were shown highest in stem and next order root and branchwood, but lowest in topwood. The differences were shown clearly between stemwood and other parts of tree, but not root and branchwood. However the significant difference is realized as most lowest value in topwood. 6. In compression strength parallel to the grain compared among the different parts of tree at the 14 percent of moisture content, highest strength was appeared in stem, next order branch and rootwood, but lowest in topwood. 7. In bending strength compared among the different parts of tree at the 14 percent of moisture content clearly highest strength was shown in branchwood, next order stem and root, but lowest in topwood. Though the branchwood has lower specific gravity than stemwood it was shown clearly high bending strength.

• Composites Research
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• v.12 no.4
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• pp.71-78
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• 1999

To determine the effect of chemical structure of linear amine curing agents on thermal and mechanical properties, standard epoxy resin DGEBA was cured with diaminodiphenyl methane (DDM), diaminodiphenyl sulphone (DDS) in a stoichiometrically equivalent ratio. From this work, the effect of aromatic amine curing agents. In contrast, the results show that the DGEBA/DDS cure system having the sulfone structure between the benzene rings had higher values in the conversion of epoxide, density, shrinkage (%), glass transition temperature, tensile modulus and strength, flexural modulus and strength than the DGEBA/DDM cure system having methylene structure between the benzene rings, whereas the DGEBA/DDM cure system presented higher values in the maximum exothermic temperature, thermal expansion coefficient, and thermal stability. These results are caused by the relative effects of sulfone group having strong electronegativity and methylene group having (+) repulsive property and stem from the effect of the conversion ratio of epoxide group. The result of fractography shows that the each grain size of the DDM/DGEBA system with feather-like structure is larger than that of the DDS/DGEBA system.