• The Korean Journal of Physiology and Pharmacology
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• 제22권5호
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• pp.555-566
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• 2018

Human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) are used in tissue repair and regeneration; however, the mechanisms involved are not well understood. We investigated the hair growth-promoting effects of hUCB-MSCs treatment to determine whether hUCB-MSCs enhance the promotion of hair growth. Furthermore, we attempted to identify the factors responsible for hair growth. The effects of hUCB-MSCs on hair growth were investigated in vivo, and hUCB-MSCs advanced anagen onset and hair follicle neogeneration. We found that hUCB-MSCs co-culture increased the viability and up-regulated hair induction-related proteins of human dermal papilla cells (hDPCs) in vitro. A growth factor antibody array revealed that secretory factors from hUCB-MSCs are related to hair growth. Insulin-like growth factor binding protein-1 (IGFBP-1) and vascular endothelial growth factor (VEGF) were increased in co-culture medium. Finally, we found that IGFBP-1, through the co-localization of an IGF-1 and IGFBP-1, had positive effects on cell viability; VEGF secretion; expression of alkaline phosphatase (ALP), CD133, and ${\beta}-catenin$; and formation of hDPCs 3D spheroids. Taken together, these data suggest that hUCB-MSCs promote hair growth via a paracrine mechanism.

• 대한소아치과학회지
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• 제41권1호
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• pp.1-7
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• 2014

가정에서 유치가 탈락하였을 경우 쉽게 구할 수 있는 저장 용액에 보관한 후, 보관되었던 유치에서 세포를 배양할 수 있다면 지금보다 더 많은 유치로부터 줄기 세포를 추출할 수 있을 것이다. 본 연구는 유치의 치수 세포가 다양한 보관 용액의 종류와 기간에 영향을 받는지 확인해 보고자 하였다. 330개의 탈락 유치를 신선군, 건조군, 세포배양액 2, 4, 7일 보관군, 우유 2, 4, 7일 보관군, 생리식염수 2, 4, 7일 보관군으로 각각 30개씩 무작위로 나누었다. 각 군 간의 유치 치수 조직 세포의 일차 배양을 시행하여 성공한 개수와 실패한 개수를 계산하여 생존 성공률을 비교하였다. 일차 배양 결과 보관 기간이 늘어날수록 일차 배양 시 세포의 생존 성공률이 낮아지는 것으로 나타났다. 2일 보관군과 4일 보관군까지 세포배양액, 우유, 생리식염수 간의 보관 용액에 따른 성공률의 유의할 만한 차이는 없었다. 그러나 7일 보관군에서는 세포배양액에 비해 우유와 생리식염수에 보관한 유치에서 성공률이 유의하게 떨어지는 것으로 나타났다. 유치 발거 후 우유나 생리식염수 등의 보관용액에 일정 기간 보관 후 그 치수 조직을 세포 배양에 사용하는 것이 가능하지만 보관 기간이 길어질수록 세포 획득 가능성이 줄어든다.

• Journal of Microbiology and Biotechnology
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• 제17권7호
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• pp.1113-1119
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• 2007

Human bone marrow-derived mesenchymal stem cells (hBMMSCs) must differentiate into osteogenic cells to allow for successful bone regeneration. In this study, we investigated the effects of different combinations of three soluble osteogenic differentiation-inducing factors [L-ascorbic acid (AC), ${\beta}$-glycerophosphate (${\beta}G$), and bone morphogenic protein-2 (BMP-2)] and the presence of a hydroxyapatite (HA) substrate on hBMMSC osteogenic differentiation in vitro. hBMMSCs were cultured in medium containing various combinations of the soluble factors on culture plates with or without HA coating. After 7 days of culture, alkaline phosphatase (ALP) activity, calcium deposition, and osteoprotegerin (OPG) and osteopontin (OPN) expression were measured. The effects of individual and combined factors were evaluated using a factorial analysis method. BMP-2 predominantly affected expression of early markers of osteogenic differentiation (ALP and OPG). HA had the highest positive effect on OPN expression and calcium deposition. The interaction between AC, ${\beta}G$, and HA had the second highest positive effect on ALP activity.

• Clinical and Experimental Reproductive Medicine
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• 제31권4호
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• pp.209-215
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• 2004

Objective: Embryonic stem (ES) cells could be differentiated into the specific cell types by alternation of culture condition and modification of gene expression. This study was performed to evaluate the differentiation protocol for mouse and human ES cells to insulin secreting cells. Methods: Undifferentiated mouse (JH-I) and human (Miz-hESI) ES cells were cultured on STO feeder layer, and embryoid bodies (EBs) were formed by suspension culture. For the differentiation, EBs were cultured by sequential system with three stage protocol. The differentiating ES cells were collected and marker gene expressions were analyzed by seIni-quantitative RT-PCR in each stage. Amount of secreted insulin levels in culture media of human ES cells were measured by human insulin specific RIA kit. Results: During the differentiation process of human ES cells, GATA-4, a-fetoprotein, glucose transporter-2 and Ngn-3 expression were increased whereas OctA was decreased progressively. Insulin and albuInin mRNAs were expressed from stage IT in mouse ES cells and from stage III in human ES cells. We detected 3.0~7.9 IlU/rnl secretion of insulin from differentiated human ES cells by in vitro culture for 36 days. Conclusion: The sequential culture system could induce the differentiation of mouse and human ES cells into insulin secreting cells. This is the fIrst report of differentiation of human ES cells into insulin secreting cells by in vitro culture with serum and insulin free medium.

• 한국수정란이식학회지
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• 제31권1호
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• pp.89-95
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• 2016

The present study was conduct to examine the $H_2O_2$ expression level and apoptosis-related gene expression levels inporcineskin-derived stem cell-like cells (pSSCs) after adipogenic, chondrogenic, and osteogenic differentiation induction. The pSSCs were obtained by digestion of porcine ear skin biopsy and cultured in each induction medium for 21 to 26 days to induce adipogenic, chondrogenic, and osteogenic differentiation, respectively. The $H_2O_2$ levels of pSSCs after induction culture were evaluated by staining with 2'7'-dichlorodihydrofluorescein diacetate ($H_2DCFDA$). The apoptotic gene expression of pSSCs after induction culture was also estimated by RT-PCR. The pSSCs have a potential to differentiate into three mesodermal cell types (adipocytes, chondrocytes, and osteoblasts). Non-induced control and chondrogenic-induced cells were showed higher $H_2DCFDA$ intensity (P<0.05) than adipogenic- and osteogenic-induced cells. The relative expression of Bax/Bcl-2 level was significantly low (P<0.05) in adipogenic- and osteogenic-induced cells compared to non-induced control. However, there was no difference in the relative expression of Bax/Bcl-2 level among differentiation induction groups. The result of the present study shows that the apoptosis of pSSCs is not detrimentally increased by differentiation induction culture, although chondrogenic-induced pSSCs showed high ROS generation level and apoptotic index similarly to those of non-induced cells.

• Bulletin of the Korean Chemical Society
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• 제32권9호
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• pp.3405-3410
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• 2011

We have investigated the cytotoxic potential of tellurium (Te) nanowires in BALB/3T3 fibroblast cells. Te nanowires were synthesized through an aqueous phase surfactant assisted method. Toxicological experiments, such as analysis of morphological changes, MTT assay, DAPI staining, and estimation of intracellular reactive oxygen species, were carried out to reveal the cytotoxic effects of Te nanowires. Te nanowires were found to be cytotoxic at all concentrations tested, in a dose-dependent manner. The UV/Vis spectra of Te nanowires suspended in a culture medium showed drastic changes and disappearance of two broad absorption peaks. The physicochemical properties such as, surface charge, size, and shape of Te nanowires were found to be altered during exposure of cells, due to the instability and agglomeration of nanowires in the culture medium. These results suggest that the chemical components of the DMEM medium significantly affect the stability of Te nanowires. In addition, TEM images revealed that necrosis was the basic pattern of cell death, which might stem from the formation of toxic moieties of tellurium, released from nanowire structures, in the bioenvironment. These observations thus suggest that Te nanomaterials may pose potential risks to environmental and human health.

• 한국임상수의학회지
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• 제40권5호
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• pp.323-329
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• 2023

Peripheral blood-derived mesenchymal stem cells (PB-MSCs) have shown promise in cell-based therapy, as they can be harvested with ease through minimally invasive procedures. This study aimed to isolate PB-MSCs from foals and mares and to compare the proliferation and cellular characteristics of the PB-MSCs between the two groups. Six pairs of mares and their foals were used in this study. MSCs were isolated from PB by direct plating in a tissue culture medium, and cell proliferation (population doubling time [PDT], and colony-forming unit-fibroblast assay [CFU-F]), and characterization (morphology, plastic adhesiveness, colony formation, trilineage differentiation) were examined. There was no significant difference in the PB-MSC yield, CFU-F, and PDT between the mares and foals. PB-MSCs from both mares and foals showed typical MSC characteristics in terms of spindle-shaped morphology, plastic adhesive properties, formation of colonies, trilineage differentiation. These results suggest that PB-MSCs isolated from horses, both adult horses, and foals, can be used for equine cell-based therapy.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2002년도 춘계학술발표대회 발표논문초록집
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• pp.20-20
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• 2002

This study was to establish the use of parthenogenetic mouse ES (P-mES02) cells as a reproducible differentiation system for mouse cardiomyocytes. To induce differentiation, P-mES02 cells were dispersed by dissociation and the formation of ES cell aggregates in differentiation medium. After 7 days in differentiation culture, the embryoid bodies (EBs) were plated onto gelatin-coated dish. Cultures were observed daily using an inverted light microscope to determine the day of contraction onset and total duration of continuous contractile activity for each contracting focus. (omitted)

• 식물조직배양학회지
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• 제24권3호
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• pp.167-173
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• 1997

The embryogenic callus (EC), from which somatic embryos could be induced, was compared with nonembryogenic callus(NE) to study the origin and features of totipotent cell in water dropwort (Oenanthe stolonifera DC). To induce and maintain of EC and the NE, meristematic stem and immature floret were inoculated in MS media supplemented with 1 mg/L 2,4-D, and with 2.5 mg/L NAA and 5mg/L BA, respectively, The EC was not induced from the NE even after subculturing in MS medium supplemented with 1 mg/L 2,4-D. Plantlets were not regenerated from the NE in hormone-free medium. In histochemical comparison of the EC with the NE by light microscopy, the EC had smaller cells in size, dense cytoplasm, and more starch granules of cells compared to the NE cells. The cell from the EC, as observed by transmission electron microscopy, had smaller vaculoes, well developed ribosomes, mitochondria, and endoplasmic reticulum, whereas the cells from the NE had larger vacuoles and underdeveloped organelles. In protein pattern from NE, EC and Somatic embryo (SE), as analyzed by SDS polyacrylamide gel electrophoresis, different proteins specific for tissue were observed: 17 and 28 KD for NE, 50, 52, 57, 66, 68 KD for EC and 20 KD for SE. DNA polymorphism was also observed between EC and NE as analyzed by RAPD (randomly amplified polymorphic DNA) method. The origin of totipotent stem cell and the relationship between irreversible genomic change arose in differentiation and the loss of totipotency in plant were discussed.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2002년도 생물공학의 동향 (X)
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• pp.289-292
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• 2002

The corneal tissue consists of three layers : epithelium, stroma, and endothelium. Central cornea is a highly differentiated tissue whereas the limbus contains the epithelial stem cell. In the present study. we report the engineering of the three-dimensional reconstructed cornea derived from rabbit limbal epithelial and stromal cells. The differentiation degree of corneal stem cells were assessed in serum concentration and inoculation density of stromal cells. Optimal condition differentiation of corneal stem cells is achieved when 5% FBS was supplemented to culture medium and $1-2{\times}10^5$ cells/ml inoculation density of stromal cells.