• Title/Summary/Keyword: stem-like cells

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Hematopoietic effect of deer antler extract fermented by Bacillus subtilis on murine marrow cells

  • Park, Yooheon;Choi, Hyeon-Son;Lee, Hyun-Sun;Suh, Hyung Joo
    • Nutrition Research and Practice
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    • v.9 no.5
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    • pp.451-458
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    • 2015
  • BACKGROUND/OBJECTIVES: We examined the chemical composition and the effect of fermented deer antler on hematopoietic factors in bone marrow cells. MATERIALS/METHODS: For the preparation of fermented deer antler extract (FAB), fermentation was carried out using Bacillus subtilis at $30^{\circ}C$ for 7 days. The hematopoietic effect of FAB was investigated hematopoietic factors in marrow cells. RESULTS: The contents of total sugar, sulfated glycosaminoglycans, and uronic acid and the dry weight gradually increased with fermentation time. The sialic acid content (from 0.14 mg/mL to 0.54 mg/mL) was the highest on the 4th day of fermentation after which it decreased. The proliferating activity of bone marrow cells increased with fermentation times. The levels of various hematopoietic growth factors were determined to verify the beneficial effect of deer antler extract fermented by B. subtilis on hematopoiesis. FAB increased the number of stem cell factors and granulocyte colony-stimulating factor in bone marrow cells. In addition, FAB augmented the burst-forming unit erythroid and total colonies in splenocyte-conditioned medium compared with non-fermented antler extract (NFA). However, FAB did not affect the mRNA levels of erythropoietin, an important factor for erythropoiesis. CONCLUSIONS: FAB, like NFA, did not directly affect hematopoiesis, but contributed to hematopoiesis by stimulating the production of hematopoietic factors.

T Cell Immune Responses against SARS-CoV-2 in the With Corona Era

  • Ji-Eun Oh
    • Biomedical Science Letters
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    • v.28 no.4
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    • pp.211-222
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    • 2022
  • After more than two years of efforts to end the corona pandemic, a gradual recovery is starting in countries with high vaccination rates. Easing public health policies for a full-fledged post-corona era, such as lifting the mandatory use of outdoor mask and quarantine measures in entry have been considered in Korea. However, the continuous emergence of new variants of SARS-CoV-2 and limitations in vaccine efficacy still remain challenging. Fortunately, T cells and memory T cells, which are key components of adaptive immunity appear to contribute substantially in COVID-19 control. SARS-CoV-2 specific CD4+/CD8+ T cells are induced by natural infection or vaccination, and rapid induction and activation of T cells is mainly associated with viral clearance and attenuated clinical severity. In addition, T cell responses induced by recognition of a wide range of epitopes were minimally affected and conserved against the highly infectious subsets of omicron variants. Polyfunctional SARS-CoV-2 specific T cell memory including stem cell-like memory T cells were also developed in COVID-19 convalescent patients, suggesting long lasting protective T cell immunity. Thus, a robust T-cell immune response appears to serve as a reliable and long-term component of host protection in the context of reduced efficacy of humoral immunity and persistent mutations and/or immune escape.

Cytotoxicity and biocompatibility of high mol% yttria containing zirconia

  • Gulsan Ara Sathi Kazi;Ryo Yamagiwa
    • Restorative Dentistry and Endodontics
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    • v.45 no.4
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    • pp.52.1-52.11
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    • 2020
  • Objectives: Yttria-stabilized tetragonal phase zirconia has been used as a dental restorative material for over a decade. While it is still the strongest and toughest ceramic, its translucency remains as a significant drawback. To overcome this, stabilizing the translucency zirconia to a significant cubic crystalline phase by increasing the yttria content to more than 8 mol% (8YTZP). However, the biocompatibility of a high amount of yttria is still an important topic that needs to be investigated. Materials and Methods: Commercially available 8YTZP plates were used. To enhance cell adhesion, proliferation, and differentiation, the surface of the 8YTZP is sequentially polished with a SiC-coated abrasive paper and surface coating with type I collagen. Fibroblast-like cells L929 used for cell adherence and cell proliferation analysis, and mouse bone marrow-derived mesenchymal stem cells (BMSC) used for cell differentiation analysis. Results: The results revealed that all samples, regardless of the surface treatment, are hydrophilic and showed a strong affinity for water. Even the cell culture results indicate that simple surface polishing and coating can affect cellular behavior by enhancing cell adhesion and proliferation. Both L929 cells and BMSC were nicely adhered to and proliferated in all conditions. Conclusions: The results demonstrate the biocompatibility of the cubic phase zirconia with 8 mol% yttria and suggest that yttria with a higher zirconia content are not toxic to the cells, support a strong adhesion of cells on their surfaces, and promote cell proliferation and differentiation. All these confirm its potential use in tissue engineering.

Human Amniotic Fluid Cells Support Expansion Culture of Human Embryonic Stem Cells (양수 세포를 이용한 인간배아줄기세포의 배양)

  • Kim, Hee-Sun;Seol, Hye-Won;Ahn, Hee-Jin;Oh, Sun-Kyung;Ku, Seung-Yup;Kim, Seok-Hyun;Choi, Young-Min;Kim, Jung-Gu;Moon, Shin-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.4
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    • pp.261-271
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    • 2004
  • Objective: This study was performed to evaluate the possibility of prolonged culture of human embryonic stem cells (hESC; SNUhES2) on human amniotic fluid cells (hAFC), which had been storaged after karyotyping. Method: The hAFC was prepared for feeder layer in the presence of Chang's medium and STO medium (90% DMEM, 10% FBS) at $37^{circ}C$ in a 5% $CO_2$ in air atmosphere. Prior to use as a feeder layer, hAFC was mitotically inactivated by mitomycin C. The hESCs on hAFC were passaged mechanically every seven days with ES culture medium (80% DMEM/F12, 20% SR, bFGF). Results: The hAFC feeder layer support the growth of undifferentiated state of SNUhES2 for at least 59 passages thus far. SNUhES2 colonies on hAFC feeder appeared slightly angular and flatter shape as compared with circular and thicker colonies observed with STO feeder layer and showed higher level with complete undifferentiation in seven days. Like hESC cultured on STO feeders, SNUhES2 grown on hAFC expressed normal karyotype, positive for alkaline phosphatase activity, high telomerase activity, Oct-4, SSEA-3, SSEA-4, Tra-1-60 and Tra-1-81 and formed embryoid bodies (EBs). Conclusion: The hAFC supports undifferentiated growth of hESC. Therefore, these results may help to provide a clinically practicable method for expansion of hESC for cell therapies.

Stem Cell Properties of Human Umbilical Cord-derived Stem Cells after Cryopreservation (냉동 보존 전후의 사람 탯줄 유래 줄기세포의 특성 분석)

  • Kang, Hyun-Mi;Park, Se-Ah;Yoon, Jin-Ah;Heo, Jin-Yeong;Kim, Hae-Kwon
    • 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.

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Differential Effect of MyD88 Signal in Donor T Cells on Graft-versus-Leukemia Effect and Graft-versus-Host Disease after Experimental Allogeneic Stem Cell Transplantation

  • Lim, Ji-Young;Ryu, Da-Bin;Lee, Sung-Eun;Park, Gyeongsin;Choi, Eun Young;Min, Chang-Ki
    • Molecules and Cells
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    • v.38 no.11
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    • pp.966-974
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    • 2015
  • Despite the presence of toll like receptor (TLR) expression in conventional $TCR{\alpha}{\beta}$ T cells, the direct role of TLR signaling via myeloid differentiation factor 88 (MyD88) within T lymphocytes on graft-versus-host disease (GVHD) and graft-versus-leukemia (GVL) effect after allogeneic stem cell transplantation (allo-SCT) remains unknown. In the allo-SCT model of C57BL/6 ($H-2^b$) ${\rightarrow}$ B6D2F1 ($H-2^{b/d}$), recipients received transplants of wild type (WT) T-cell-depleted (TCD) bone marrow (BM) and splenic T cells from either WT or MyD88 deficient (MyD88KO) donors. Host-type ($H-2^d$) P815 mastocytoma or L1210 leukemia cells were injected either subcutaneously or intravenously to generate a GVHD/GVL model. Allogeneic recipients of MyD88KO T cells demonstrated a greater tumor growth without attenuation of GVHD severity. Moreover, GVHD-induced GVL effect, caused by increasing the conditioning intensity was also not observed in the recipients of MyD88KO T cells. In vitro, the absence of MyD88 in T cells resulted in defective cytolytic activity to tumor targets with reduced ability to produce IFN-${\gamma}$ or granzyme B, which are known to critical for the GVL effect. However, donor T cell expansion with effector and memory T-cell differentiation were more enhanced in GVHD hosts of MyD88KO T cells. Recipients of MyD88KO T cells experienced greater expansion of Foxp3- and IL4-expressing T cells with reduced INF-${\gamma}$ producing T cells in the spleen and tumor-draining lymph nodes early after transplantation. Taken together, these results highlight a differential role for MyD88 deficiency on donor T-cells, with decreased GVL effect without attenuation of the GVHD severity after experimental allo-SCT.

Morphological Characteristics of Growth of Rice and Barnyardgrass under Various Cropping Patterns - III. Difference in Stem Surface and Ultrastructures (재배양식(栽培樣式)에 따른 벼와 피의 생장(生長) 및 해부형태학적(解剖形態學的) 차이(差異) - III. 재배양식(栽培樣式)에 따른 벼와 피간(間의) 미세구조(微細構造) 차이(差異))

  • Chon, S.U.;Guh, J.O.;Lee, Y.M.
    • Korean Journal of Weed Science
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    • v.14 no.2
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    • pp.144-155
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    • 1994
  • Scanning electron micrography and Transmission electron micrography studies were performed on the surfaces of stem(coleoptiles) of rice and barnyardgrass under various cropping patterns at 5 days after seeding or transplanting. Stem surfaces of rice under dry condition possessed rodlet-like epicuticular waxes, trichomes, stomata and cork cell layer in both broadcast and drilled rice while barnyardgrass had only thread-like epicuticular waxes, but trichomes, stomata and cork cell layers were not found. However, rice under water condition has no rodlet-like waxes, stomata and cork cells, and barnyardgrass was similar to that under dry condition. On the other hand, those on stems of 8-day and 25-day transplanted rice were well arranged and developed compared with those of direct seeded rice. Electron micrography showed significant difference in thickness of outer epidermal cell walls of rice and barnyardgrass under dry, water and transplanting condition. Thickness of outer epidermal cell was greater in transplanted rice, dry direct seeded rice, water-direct seeded rice and barnyardgrass in descending order.

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Evaluation of apoptosis after ionizing radiation in feeding and starving rats

  • Lee, Jae-Hyun;Cho, Kyung-Ja;Hong, Seok-Il;Park, Min-Kyung
    • Korean Journal of Veterinary Pathology
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    • v.2 no.1
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    • pp.37-46
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    • 1998
  • It has been known that $\gamma$-irradiation usually induces cell death in regenerating stem cell in normal tissues like skin, intestine and hematopoietic organ. The experiment were carried out to evaluate the early response of radiation injury in radiosensitive and intermediate radiosensitive tissues in feeding and starving rats with the doses of 3.5 and 7.0 Gy. The results of the study showed that the histological phenomenon was apoptosis in the doses of the radiation as the early response of tissue injury. Apoptosis were showed organ-specific and cellular specific responses suggesting that the selection of apoptosis be exactly focused on highly renewal organs and cells. It was interesting that the rats starved for 72 hours prior to irradiation induced less apoptosis in liver than fed rats. As for cellular responses it appeared that apoptotic cells were mostly distributed in ductal or periportal cells in liver of feeding rats unlikely in liver of Starving rots which showed no difference in zonal distribution. In salivary gland apoptotic cells in fed rats were highly induced in intercalating and ductal cell population than in acinar cell population although unlikely in starved rats. This study showed the value of apoptosis using the detection system of TUNEL for evaluating cellular damage after radiation injury and the diminished effect of starvation on cell damage after ionizing irradiation.

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The Decreased Expression of Fbxw7 E3 Ligase Mediated by Cancer Upregulated Gene 2 Confers Cancer Stem Cell-like Phenotypes (CUG2 유전자에 의하여 감소된 FBXW7 E3 ligase 발현이 유사-종양줄기세포 표현형을 유도)

  • Yawut, Natpaphan;Kim, Namuk;Budluang, Phatcharaporn;Cho, Il-Rae;Kaowinn, Sirichat;Koh, Sang Seok;Kang, Ho Young;Chung, Young-Hwa
    • Journal of Life Science
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    • v.32 no.4
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    • pp.271-278
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    • 2022
  • The detailed mechanism by which cancer upregulated gene 2 (CUG2) overexpression induces cancer stem cell-like phenotypes is not fully understood. The downregulation of FBXW7 E3 ligase, a tumor suppressor known for its proteolytic regulation of oncogenic proteins such as cyclin E, c-Myc, Notch, and Yap1, has been frequently reported in several types of tumor tissues, including those in the large intestine, cervix, and stomach. Therefore, we investigated whether FBXW7 is involved in CUG2-induced oncogenesis. In this study, the decreased expression of FBXW7 was examined in human lung adenocarcinoma A549 (A549-CUG2) and human bronchial BEAS-2B cells (BEAS-CUG2) overexpressing CUG2 and compared with control cells stably expressing an empty vector (A549-Vec or BEAS-Vec). Treatment with MG132 (a proteosome inhibitor) prevented the degradation of FBXW7 and Yap1 proteins, which are substrates of the FBXW7 E3 ligase. To address the role of Fbxw7 in the development of cancer stem cell (CSC) phenotypes, we suppressed Fbxw7 protein levels using its siRNA. We observed that decreased levels of FBXW7 enhanced cell migration, invasion, and spheroid size and number in A549-Vec and BEAS-Vec cells. The enforced expression of FBXW7 produced the opposite results in A549-CUG2 and BEAS-CUG2 cells. Furthermore, the downregulation of FBXW7 elevated the activities of EGFR, Akt, and ERK1/2 and upregulated β-catenin, Yap1, and NEK2, while the enforced expression of FBXW7 generated the opposite results. We thus propose that FBXW7 downregulation induced by CUG2 confers CSC-like phenotypes through the upregulation of both the EGFR-ERK1/2 and β-catenin-Yap1-NEK2 signaling pathways.

Glucose Controls the Expression of Polypyrimidine Tract-Binding Protein 1 via the Insulin Receptor Signaling Pathway in Pancreatic β Cells

  • Jeong, Da Eun;Heo, Sungeun;Han, Ji Hye;Lee, Eun-young;Kulkarni, Rohit N.;Kim, Wook
    • Molecules and Cells
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    • v.41 no.10
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    • pp.909-916
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    • 2018
  • In pancreatic ${\beta}$ cells, glucose stimulates the biosynthesis of insulin at transcriptional and post-transcriptional levels. The RNA-binding protein, polypyrimidine tract-binding protein 1 (PTBP1), also named hnRNP I, acts as a critical mediator of insulin biosynthesis through binding to the pyrimidine-rich region in the 3'-untranslated region (UTR) of insulin mRNA. However, the underlying mechanism that regulates its expression in ${\beta}$ cells is unclear. Here, we report that glucose induces the expression of PTBP1 via the insulin receptor (IR) signaling pathway in ${\beta}$ cells. PTBP1 is present in ${\beta}$ cells of both mouse and monkey, where its levels are increased by glucose and insulin, but not by insulin-like growth factor 1. PTBP1 levels in immortalized ${\beta}$ cells established from wild-type (${\beta}IRWT$) mice are higher than levels in ${\beta}$ cells established from IR-null (${\beta}IRKO$) mice, and ectopic re-expression of IR-WT in ${\beta}IRKO$ cells restored PTBP1 levels. However, PTBP1 levels were not altered in ${\beta}IRKO$ cells transfected with IR-3YA, in which the Tyr1158/1162/1163 residues are substituted with Ala. Consistently, treatment with glucose or insulin elevated PTBP1 levels in ${\beta}IRWT$ cells, but not in ${\beta}IRKO$ cells. In addition, silencing Akt significantly lowered PTBP1 levels. Thus, our results identify insulin as a pivotal mediator of glucose-induced PTBP1 expression in pancreatic ${\beta}$ cells.