• Title/Summary/Keyword: Stem number

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Comparison of Neural Cell Differentiation of Human Adipose Mesenchymal Stem Cells Derived from Young and Old Ages (연령별 지방 중간엽 유래 줄기세포의 신경세포로의 분화 능력 비교)

  • Jo, Jung-Youn;Kang, Sung-Keun;Choi, In-Su;Ra, Jeong-Chan
    • Development and Reproduction
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    • v.13 no.4
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    • pp.227-237
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    • 2009
  • Recently, adipose mesenchymal stem cells (AdMSC) that are similar to bone marrow MSC and blood derived MSC are thought to be another source for stem cell therapy. However, the diseases that can be applied for stem cells therapy are age-dependent degenerative diseases. Accordingly, the present study investigated the growth and differentiation potential to neural cells of human AdMSC (hAdMSC) obtained from aged thirty, forty and fifty. The growth of cells and cell viability were measured by passage and neural differentiation of hAdMSC was induced in neural differentiation condition for 10 days. Our results demonstrated that cell number, viability and morphology were not different from hAdMSC by age and passage. Immunofluorescence analysis of neural cell marker (TuJ1, NSE, Sox2, GFAP or MAP2) demonstrated no significant differences in neural cell differentiation by age and passage. As the number of passage was increased, the mRNA level of MAP2 and Sox2 was decreased in hAdMSC from age of 50 compared to hAdMSC from age of 30. In conclusion, the present study demonstrated that ability of neural cell differentiation of hAdMSC was maintained with ages, suggesting that autologous stem cells from aged people can be applied for stem cell therapy with age-dependent neural disease with the same stem cell quality and ability as stem cell derived from young age.

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A Study on Direct Sowing Culture of Scutellaria baicalensis GEORGE Cultivated after Barley (황금의 맥후작 직파 재배 연구)

  • 권병선;신종섭
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2001.11b
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    • pp.49-61
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    • 2001
  • This study was carried out to determine the effect of various fertilizer levels, sowing time and planting density on the growth and yield of Scuteliaria baicalensis GEORGE cultivated after the barley in the southern coastal areas of Korea under the non- mulching condition by direct sowing culture. The flowering date of medium dressing plot(N : P$_2$O$\sub$5/ : K$_2$O=9 : 13.5 : 9kg/10a) and heavy dression plot(N : P$_2$O$\sub$5/, : K$_2$O=12 :18 : 12kg/10a) were July 23. The flowering date of the medium and the heavy dressing plot was delayed by 3days compared with that of non-fertilizing plot. The growth characteristics such as stem length, diameter of main stem, number of branch per plant, main root length, main root length, main root thickness and dry weight of stem leaves were more increased at medium dressing plot than that of other fertilizer levels, The root dry weight of in Scutellaria baicalensis GEORGE cultivated after barley was hlghest at the fertilizing plot of N : P$_2$O$\sub$5/ : K$_2$O=9 : 13.5 : 9kg/10a. The dried-root yield was 178kg in medium dressing plot, 167kg in standard dressing pot, and 126kg in non-dressing plot, The dried-root yield of medium dressing plot was 7% and 41 cie higher than that of standard dressing pot and non-dressing control plot, respectively. Emergence and flowering dates in the sowing time of June 1 were earlier than those of the other sowing times. In the sowing time of June 1, length and diameter of main stem, number of node per main stem, number of branch per plant and dry weight of stem leaves were greater than those of sowing times of June 10 and June 20. Yield components such as main stem length and diameter, main stem numbers, branches per plant, dry weight of stem leaves, main root length and thickness, number of large root and fine root per plant, and dry weight of root were the highest at the sewing time of June 1 as the yield of 71.3kg/10a. Optimum sowing time of Scuteilaria baicaiensis GEORGE cultivated after barley was June 1 in southern areas of Korea. Stem length was long in dense planting of 20${\times}$10cm and short in spacious planting of 30${\times}$10cm and 40${\times}$10cm by direct sowing cultivated after barley. Stem diameter was thick in spacious planting of 30${\times}$10cm and 40${\times}$10cm and was thin in dense planting of 20${\times}$10cm by direct sowing cultivated after barley. Length and dry weight of root per plant were decreased in dense planting of 20${\times}$10cm and were increased in spacious planting of 30${\times}$10cm and 40${\times}$10cm by direct sowing cultivated after barley. Yield of dry root was highest in optimum planting density(30${\times}$10cm 33 plants/㎡) by direct sowing cultivated after barley. The correlation coefficient between number of planting plant and stem length showed highly positive correlation. These characters of stem diameter, number of branches, main root length and yield of dry root mentioned above showed negative correlations with planting plants.

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Long-term Cryopreservation of Mesenchymal Stem Cells Derived from Human Eyelid Adipose and Amniotic Membrane: Maintenance of Stem Cell Characteristics

  • Song, Yeon-Hwa;Park, Se-Ah;Yun, Su-Jin;Yang, Hye-Jin;Yoon, A-Young;Kim, Haek-Won
    • Development and Reproduction
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    • v.15 no.4
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    • pp.339-347
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    • 2011
  • Human eyelid adipose-derived stem cells (hEAs) and amniotic mesenchymal stem cells (hAMs) are very valuable sources for the cell therapeutics. Both types of cells have a great proliferating ability in vitro and a multipotency to differentiate into adipocytes, osteoblasts and chondrocytes. In the present study, we evaluated their stem cell characteristics after long-time cryopreservation for 6, 12 and 24 months. When frozen-thawed cells were cultivated in vitro, their cumulative cell number and doubling time were similar to freshly prepared cells. Also they expressed stem cell-related genes of SCF, NANOG, OCT4, and TERT, ectoderm-related genes of NCAM and FGF5, mesoderm/endoderm-related genes of CK18 and VIM, and immune-related genes of HLA-ABC and ${\beta}$2M. Following differentiation culture in appropriate culture media for 2-3 weeks, both types of cells exhibited well differentiation into adipocyte, osteoblast, and chondrocyte, as revealed by adipogenic, osteogenic or chondrogenic-specific staining and related genes, respectively. In conclusion, even after long-term storage hEAs and hAMs could maintain their stem cell characteristics, suggesting that they might be suitable for clinical application based on stem cell therapy.

Neurogenic potentials of human amniotic fluid-derived stem cells according to expression levels of stem cell markers and ingredients of induction medium

  • Lim, Eun Hye;Cho, Jung Ah;Park, Ho;Song, Tae Jong;Kim, Woo Young;Kim, Kye Hyun;Lee, Kyo Won
    • Journal of Genetic Medicine
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    • v.12 no.1
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    • pp.31-37
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    • 2015
  • Purpose: We investigated the neurogenic potentials of amniotic fluid-derived stem cells (AFSCs) according to the expression levels of stem cell markers and ingredients in the neural induction media. Materials and Methods: Four samples of AFSCs with different levels of Oct-4 and c-kit expression were differentiated neurally, using three kinds of induction media containing retinoic acid (RA) and/or a mixture of 3-isobutyl-1-methylxanthine/indomethacin/insulin (neuromix), and examined by immunofluorescence and reverse transcription-polymerase chain reaction (RT-PCR) for their expression of neurospecific markers. Results: The cells in neuromix-containing media displayed small nuclei and long processes that were characteristic of neural cells. RT-PCR analysis revealed that the number of neural markers showing upregulation was greater in cells cultured in the neuromix-containing media than in those cultured in RA-only medium. Neurospecific gene expression was also higher in Oct-4 and c-kit double-positive cells than in c-kit-low or -negative cells. Conclusion: The stem cell marker c-kit (rather than Oct-4) and the ingredient neuromix (rather than RA) exert greater effects on neurogenesis of AFSCs.

Influence of Sowing Dates and Nursery Period on Growth and Yield of Alisma plantago L. in Mono Cropping (파종기 및 육묘기간이 단작택사의 생육 및 수량에 미치는 영향)

  • 장영석;최달호;권병선;신종섭
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2000.05a
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    • pp.38-44
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    • 2000
  • To determine the optimum sowing date and nursery period in Alisma piantago in the southern area of Korea, Alisma plantago local cultivar was grown under three different sowing dates and nursery periods. The plant height, leaf width and leaf length at the area sown on the seedbed in June 30 and conducted the growing seedling in 30 days is 12.1cm, 2.5cm and 3.1cm respectively and it shows tess decrease than that of plant height, leaf width and leaf length with 12.3cm, 2.6cm and 3.2cm respectively which was sown on the seedbed in June 20 and carried out the growing seedling in 30 days, and it also shows less decrease than that of plant height, leaf width and leaf length with 15.2cm, 3.1cm and 5.2cm respectively at the area sown on the seedbed in July 10 and condected the growing seedling in the same date. The stem length, number of stems and diameter of root at the area sown on the seedbed in June 30 and conducted the growing seedling in 30 days is 31cm, 11 and 3.7cm respectively and it shows more increase than that of stem length, number of stem and diameter of and on the other hand shows tess decrease than that of stem length, number of stems and diameter of root with 30cm, 10cm, 35cm respectively which was sown on the seedbed in 30 days, and on the other hand shows less decrease than that of stem length, number of stems and diameter of root with 32cm,13cm,3.9cm respectively at the area sown on the seedbed in July 10 and conducted the growing seedling in the same date, The fresh yield of roots at the area sown on the seedbed in June 30 and conducted the growing seedling in 30 days is 431.4Kg/10a and it shows more increase of 26.9Kg than that of fresh yield of root with 410.59g110a which was sown on the seedbed in lune 20an4 carried out the growing seedling in 30 days, and it also shows more increase of 7Kg than that of fresh yield of root with 430.4Kg/10a at the area sown on the seedbed in July 10 and conducted the growing seedling in the same date. The dry yield of roots at the area on the seedbed in June 30 and conducted the growing seedling in 30 days is 351.9Kg110a which was sown on the seedbed in June 20 and carried out the growing seedling in 30 days, and it also shows more increase of 16.4Kg than that of dry yield of root with 335.4Kg110a at the area sown on the seedbed in July 10 and conducted the growing seedling in the same date. Therefore, the seedling period of proper seedbed for high yield bumper crop of Alisma plantago for exporting to Japan is June 30 and the number of days for seedling is 30 days.

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Forced Expression of HoxB4 Enhances Hematopoietic Differentiation by Human Embryonic Stem Cells

  • Lee, Gab Sang;Kim, Byung Soo;Sheih, Jae-hung;Moore, Malcolm AS
    • Molecules and Cells
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    • v.25 no.4
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    • pp.487-493
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    • 2008
  • HoxB4 has been shown to enhance hematopoietic engraftment by hematopoietic stem cells (HSC) from differentiating mouse embryonic stem cell (mESC) cultures. Here we examined the effect of ectopic expression of HoxB4 in differentiated human embryonic stem cells (hESCs). Stable HoxB4-expressing hESCs were established by lentiviral transduction, and the forced expression of HoxB4 did not affect stem cell features. HoxB4-expressing hESC-derived CD34+ cells generated higher numbers of erythroid and blast-like colonies than controls. The number of CD34+ cells increased but CD45+ and KDR+ cell numbers were not significantly affected. When the hESC derived CD34+ cells were transplanted into $NOD/SCID{\beta}2m-/-$ mice, the ectopic expression of HoxB4 did not alter their repopulating capacity. Our findings show that overexpression of HoxB4 in differentiating hESCs increases hematopoietic colony formation and hematopoietic cell formation in vitro, but does not affect in vivo repopulation in adult mice hosts.

Combination stem cell therapy using dental pulp stem cells and human umbilical vein endothelial cells for critical hindlimb ischemia

  • Kim, Chung Kwon;Hwang, Ji-Yoon;Hong, Tae Hee;Lee, Du Man;Lee, Kyunghoon;Nam, Hyun;Joo, Kyeung Min
    • BMB Reports
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    • v.55 no.7
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    • pp.336-341
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    • 2022
  • Narrowing of arteries supplying blood to the limbs provokes critical hindlimb ischemia (CLI). Although CLI results in irreversible sequelae, such as amputation, few therapeutic options induce the formation of new functional blood vessels. Based on the proangiogenic potentials of stem cells, in this study, it was examined whether a combination of dental pulp stem cells (DPSCs) and human umbilical vein endothelial cells (HUVECs) could result in enhanced therapeutic effects of stem cells for CLI compared with those of DPSCs or HUVECs alone. The DPSCs+ HUVECs combination therapy resulted in significantly higher blood flow and lower ischemia damage than DPSCs or HUVECs alone. The improved therapeutic effects in the DPSCs+ HUVECs group were accompanied by a significantly higher number of microvessels in the ischemic tissue than in the other groups. In vitro proliferation and tube formation assay showed that VEGF in the conditioned media of DPSCs induced proliferation and vessel-like tube formation of HUVECs. Altogether, our results demonstrated that the combination of DPSCs and HUVECs had significantly better therapeutic effects on CLI via VEGF-mediated crosstalk. This combinational strategy could be used to develop novel clinical protocols for CLI proangiogenic regenerative treatments.

Development of an effective dissociation protocol for isolating mesenchymal stem cells from bovine intermuscular adipose tissues

  • Jeong Min Lee;Hyun Lee;Seung Tae Lee
    • Journal of Animal Reproduction and Biotechnology
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    • v.38 no.1
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    • pp.10-16
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    • 2023
  • Intermuscular fat is essential for enhancing the flavor and texture of cultured meat. Mesenchymal stem cells derived from intermuscular adipose tissues are a source of intermuscular fat. Therefore, as a step towards developing a platform to derive intermuscular fat from mesenchymal stem cells (MSCs) for insertion between myofibrils in cultured beef, an advanced protocol of intermuscular adipose tissue dissociation effective to the isolation of MSCs from intermuscular adipose tissues was developed in cattle. To accomplish this, physical steps were added to the enzymatic dissociation of intermuscular adipose tissues, and the MSCs were established from primary cells dissociated with physical step-free and step-added enzymatic dissociation protocols. The application of a physical step (intensive shaking up) at 5 minutes intervals during enzymatic dissociation resulted in the greatest number of primary cells derived from intermuscular adipose tissues, showed effective formation of colony forming units-fibroblasts (CFU-Fs) from the retrieved primary cells, and generated MSCs with no increase in doubling time. Thus, this protocol will contribute to the stable supply of good quality adipose-derived mesenchymal stem cells (ADMSCs) as a fat source for the production of marbled cultured beef.

The Differentiation of Pluripotent Stem Cells towards Endothelial Progenitor Cells - Potential Application in Pulmonary Arterial Hypertension

  • Kezhou Qin;Jia Lei;Jun Yang
    • International Journal of Stem Cells
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    • v.15 no.2
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    • pp.122-135
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    • 2022
  • Background and Objectives: Endothelial progenitor cells (EPCs) and endothelial cells (ECs) have been applied in the clinic to treat pulmonary arterial hypertension (PAH), a disease characterized by disordered pulmonary vasculature. However, the lack of sufficient transplantable cells before the deterioration of disease condition is a current limitation to apply cell therapy in patients. It is necessary to differentiate pluripotent stem cells (PSCs) into EPCs and identify their characteristics. Methods and Results: Comparing previously reported methods of human PSCs-derived ECs, we optimized a highly efficient differentiation protocol to obtain cells that match the phenotype of isolated EPCs from healthy donors. The protocol is compatible with chemically defined medium (CDM), it could produce a large number of clinically applicable cells with low cost. Moreover, we also found PSCs-derived EPCs express CD133, have some characteristics of mesenchymal stem cells and are capable of homing to repair blood vessels in zebrafish xenograft assays. In addition, we further revealed that IPAH PSCs-derived EPCs have higher expression of proliferation-related genes and lower expression of immune-related genes than normal EPCs and PSCs-derived EPCs through microarray analysis. Conclusions: In conclusion, we optimized a highly efficient differentiation protocol to obtain PSCs-derived EPCs with the phenotypic and molecular characteristics of EPCs from healthy donors which distinguished them from EPCs from PAH.

Growth Characteristics of Small and Medium Type Watermelon According to Number of Stem Training and Position of Fruit Setting in the Winter Season (겨울철 줄기유인 수 및 착과 위치에 따른 중·소과종 수박의 생육 특성)

  • Kim, So-Hui;Choi, Gyeong-Lee;Choi, Su-Hyun;Lim, Mi-Young;Jeong, Ho-Jeong
    • Journal of Bio-Environment Control
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    • v.29 no.2
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    • pp.189-195
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    • 2020
  • This research was carried out to find the proper number of stem training and position of fruit setting that can be stably produced for the cultivation in small and medium types of watermelon during winter. The treatments for the number of stem training were 2-, 3-, 4-stems, respectively. Growth characteristics (plant height, stem diameter, no. of node, etc.) by number of stem training were higher in 2-stem than in 3-4-stem. However, Fruit characteristics such as weight, length, width were high in the 4-stem. There is no significant difference between the soluble solids and fruit setting rate depending on the stem training. The position of fruit setting were three points: 2nd, 3rd, 4th female flower positions. The fruit setting is one fruit per plant. The average fruit setting nodes of 2nd, 3rd and 4th female flowers were 11.5, 15.8 and 23.1 nodes, respectively. The 4th female flower was 0.8 kg heavier than 2nd female flower because of its increased weight as position of fruit setting was higher. However, the soluble solids decreased as the position of fruit setting increased, with the second female flower being 1.3°Bx higher than the 4th female flower. The Fruit setting rate was no significant difference. Considering the growth and fruit characteristics, it is believed that the small and medium-sized watermelon in winter will have a high quality production of watermelon when the stem training is 3-stem and the position of fruit setting is 3rd female flower. However, it is thought that additional studies are needed to stabilize the income of watermelon-growing farms, such as planting distance and adhesion of small and medium-sized varieties.