• Title/Summary/Keyword: multiple stem

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Dysfunctional pancreatic cells differentiated from induced pluripotent stem cells with mitochondrial DNA mutations

  • So, Seongjun;Lee, Song;Lee, Yeonmi;Han, Jongsuk;Kang, Soonsuk;Choi, Jiwan;Kim, Bitnara;Kim, Deokhoon;Yoo, Hyun-Ju;Shim, In-Kyong;Oh, Ju-Yun;Lee, Yu-Na;Kim, Song-Cheol;Kang, Eunju
    • BMB Reports
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    • v.55 no.9
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    • pp.453-458
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    • 2022
  • Diabetes mellitus (DM) is a serious disease in which blood sugar levels rise abnormally because of failed insulin production or decreased insulin sensitivity. Although many studies are being conducted for the treatment or early diagnosis of DM, it is not fully understood how mitochondrial genome (mtDNA) abnormalities appear in patients with DM. Here, we induced iPSCs from fibroblasts, PBMCs, or pancreatic cells of three patients with type 2 DM (T2D) and three patients with non-diabetes counterpart. The mtDNA mutations were detected randomly without any tendency among tissues or patients. In T2D patients, 62% (21/34) of iPSC clones harbored multiple mtDNA mutations, of which 37% were homoplasmy at the 100% mutation level compared to only 8% in non-diabetes. We next selected iPSC clones that were a wild type or carried mutations and differentiated into pancreatic cells. Oxygen consumption rates were significantly lower in cells carrying mutant mtDNA. Additionally, the mutant cells exhibited decreased production of insulin and reduced secretion of insulin in response to glucose. Overall, the results suggest that screening mtDNA mutations in iPSCs from patients with T2D is an essential step before pancreatic cell differentiation for disease modeling or autologous cell therapy.

Retrovirus Vector-Mediated Gene Transfer to the Chicken Blastodermal Cells Cultured In Vitro (체외 배양된 닭 배반엽 세포에 대한 Retrovirus Vector를 이용한 유전자 전이)

  • Park, Sung-Joon;Koo, Bon-Chul;Kwon, Mo-Sun;Chae, Whi-Gun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.257-262
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    • 2010
  • The purpose of this study is to establish a basic culture system enabling in vitro culture of chicken blastodermal cells and to test the feasibility of retrovirus-mediated gene transfer to the cultured cells. The blastodermal cells were isolated from freshly laid eggs of stage X and cultured with or without STO feeder layer cells. Stem cell-like morphology was maintained after multiple passages and RT-PCR analysis proved expression of several stem cell specific genes. Immunocytochemical analysis using antibodies of anti-EMA-1 and anti-SSEA-1 also showed the feature of stem cells. Infection of the cultured blastodermal cells with LNCGW retrovirus vector resulted in successful transfer of foreign genes. The results of this study may be useful in establishing stem cell-mediated transgenic chicken production.

A Simple Method for the Preparation of Crude Gintonin from Ginseng Root, Stem, and Leaf

  • Pyo, Mi-Kyung;Choi, Sun-Hye;Shin, Tae-Joon;Hwang, Sung-Hee;Lee, Byung-Hwan;Kang, Ji-Yeon;Kim, Hyeon-Joong;Lee, Soo-Han;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.35 no.2
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    • pp.209-218
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    • 2011
  • Ginseng has been used as a general tonic agent to invigorate the human body as an adaptogenic agent. In a previous report, we have shown that ginseng contains a novel glycolipoprotein called gintonin. The main function of gintonin is to transiently enhance intracellular free $Ca^{2+}$ $[Ca^{2+}]_i$ levels in animal cells. The previous method for gintonin isolation included multiple steps using organic solvents. In the present report, we developed a simple method for the preparation of crude gintonin from ginseng root as well as stem and leaf, which produced a higher yield of gintonin than the previous one. The yield of gintonin was 0.20%, 0.29%, and 0.81% from ginseng root, stem, and leaf, respectively. The apparent molecular weight of gintonin isolated from stem and leaf through sodium dodecyl sulfate polyacrylamide gel electrophoresis was almost same as that from root but the compositions of amino acids, carbohydrates or lipids differed slightly between them. We also examined the effects of crude gintonin from ginseng root, stem, and leaf on endogenous $Ca^{2+}$-activated $Cl^-$ channel (CaCC) activity of Xenopus oocytes through mobilization of $[Ca^{2+}]_i$. We found that the order of potency for the activation of CaCC was ginseng root > stem > leaf. The $ED_{50}$ was $1.4{\pm}1.4$, $4.5{\pm}5.9$, and $3.9{\pm}1.1$ mg/mL for root, stem and leaf, respectively. In the present study, we demonstrated for the first time that in addition to ginseng root, ginseng stem and leaf also contain gintonin. Gintonin can be prepared from a simple method with higher yield of gintonin from ginseng root, stem, and leaf. Finally, these results demonstrate the possibility that ginseng stem and leaf could also be utilized for ginstonin preparation after a simple procedure, rather than being discarded.

Calibration Update for the Measuring Total Nitrogen Content in Rice Plant Tissue Using the Near Infrared Spectroscopy

  • Kwon, Young-Rip;Song, Young-Eun;Choi, Dong-Chil;Ryu, Jeong
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.54 no.1
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    • pp.29-35
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    • 2009
  • The aim of the present study was to update the calibration that is used for the measurement of the total nitrogen content in the rice plant samples by using the visible and near infrared spectrum. Before the equation merge, correlation coefficient of calibration equation for nitrogen content on each rice parts was 0.945 (Leaf), 0.928 (Stem), and 0.864 (Whole plant), respectively. In the calibration models created by each part in the rice plant under the various regression method, the calibration model for the leaf was recorded with relatively high accuracy. Among of those, the calibration equation developed by Partial least squares (PLS) method was more accurate than the Multiple linear regression (MLR) method. The calibration equation was sensitive based on variety and location variations. However, we have merged and enlarged various of the samples that made not only to measure the nitrogen content more accurately, but also later sampling populations became more diversified. After merging, $R^2$ value becomes more accurate and significantly to 0.950 (L.), 0.974 (S.), 0.940 (W.). Also, after removal of outlier, R2 values increased into 0.998, 0.995, and 0.997. In view of the results so far achieved, Standard error of prediction (SEP) and SEP (C) were reduced in the stem and whole plant. Biases were reduced in the leaf, stem as well as whole plant. Slopes were high in the stem. Standard deviation reduced in the stem but $R^2$ was high in the stem and whole plant. Result was indicated that calibration equation make update, and updating robust calibration equation from merge function and multi-variate calibration.

The effect of water status on productive and flowering variables in young 'Arbequina' olive trees under limited irrigation water availability in a semiarid region of Chile

  • Beya-Marshall, Victor;Herrera, Julio;Fichet, Thomas;Trentacoste, Eduardo R.;Kremer, Cristian
    • Horticulture, Environment, and Biotechnology : HEB
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    • v.59 no.6
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    • pp.815-826
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    • 2018
  • The intense drought affecting olive production in Northern Chile underscores the need to research non-traditional irrigation strategies to obtain the best crop performance. Accordingly, this study aimed to obtain preliminary data to guide future research on this topic. Different water replenishment levels on crop evapotranspiration ($ET_c$ ; 13.5, 27.0, 40.5, and 54%) were established in a young orchard, cv. Arbequina, from the end of fruit drop (EFD) to full bloom in the next season. We evaluated the influence of plant water status (${\Psi}_{stem}$ ) and crop load, considered as function of fruit number divided by trunk cross-sectional area, on reproductive and productive variables using multiple linear regressions. Our results show that crop load and ${\Psi}_{stem}$ measured from EFD to harvest affected yield components. Nevertheless, ${\Psi}_{stem}$ had the strongest influence on fruit size, pulp development, oil accumulation, and yield. Oil content and yield were reduced by 54% and 50% for each MPa, respectively, from ${\Psi}_{stem\;EFD-H}$ -1.8 MPa, an effect that intensified as crop load increased. During the period of flower development (September-November), the number of flowers per inflorescence and percentage of perfect flowers were reduced when ${\Psi}_{stem}$ was less than -2.0 MPa. These preliminary results showed that bud differentiation, inflorescence and flower formation are highly sensitive to water deficit.

Optimization of Culture Conditions for Maintaining Pig Muscle Stem Cells In Vitro

  • Choi, Kwang-Hwan;Yoon, Ji Won;Kim, Minsu;Jeong, Jinsol;Ryu, Minkyung;Park, Sungkwon;Jo, Cheorun;Lee, Chang-Kyu
    • Food Science of Animal Resources
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    • v.40 no.4
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    • pp.659-667
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    • 2020
  • Muscle stem cells isolated from domestic animals, including cows and pigs, were recently spotlighted as candidates for the production of alternative protein resources, so-called cultured meat or lab-grown meat. In the present study, we aimed to optimize the in vitro culture conditions for the long-term expansion of pig muscle stem cells via the screening of various signaling molecules. Pig muscle stem cells were collected from the biceps femoris muscles of 3-d-old crossbred pigs (Landrace×Yorkshire×Duroc, LYD) and cultured in minimum essential medium-based growth media. However, the pig muscle stem cells gradually lost their proliferation ability and featured morphologies during the long-term culture over two weeks. To find suitable in vitro culture conditions for an extended period, skeletal muscle growth medium-2, including epidermal growth factor (EGF), dexamethasone, and a p38 inhibitor (SB203580), was used to support the stemness of the pig muscle stem cells. Interestingly, pig muscle stem cells were stably maintained in a long-term culture without loss of the expression of myogenic marker genes as determined by PCR analysis. Immunostaining analysis showed that the stem cells were capable of myogenic differentiation after multiple passaging. Therefore, we found that basal culture conditions containing EGF, dexamethasone, and a p38 inhibitor were suitable for maintaining pig muscle stem cells during expanded culture in vitro. This culture method may be applied for the production of cultured meat and further basic research on muscle development in the pig.

A Case Report of Peripheral Neuropathy after Induction Chemotherapy before Autologous Peripheral Blood Stem Cell Transplantation in a Patient with Multiple Myeloma with Amyloidosis Treated with Complex Korean Medicine Treatment (아밀로이드증을 동반한 다발골수종 환자의 자가말초혈액조혈모세포 이식 전 유도항암화학요법 후 발생한 말초신경병증에 대한 복합 한의 치험 1례)

  • Mariah Kim;Seyeon Lee;Kibeom Ku;Irang Nam;Minhwa Kim;So-yeon Kim
    • The Journal of Internal Korean Medicine
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    • v.44 no.5
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    • pp.1092-1100
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    • 2023
  • Introduction: We present a case of multiple myeloma with amyloidosis, which has features of peripheral neuropathy after induction chemotherapy before autologous peripheral blood stem cell transplantation, in a 56-year-old woman with Korean medicine. Case Presentation: For 17 days of hospitalization, the patient with complaints of numbness and a tingling sensation in the hands and feet was treated with acupuncture, herbal medicine. To reduce the symptoms, we provided Korean medicine treatments, including herbal medicine, acupuncture, and moxibustion. The Visual Analog Scale (VAS) was used to evaluate the results of the treatment. Until discharge, the VAS scores decreased for both hands and the foot tingling sensation. Conclusion: According to these results, Korean medicine treatment may be considered an effective treatment for tingling sensations in a patient with multiple myeloma with amyloidosis. Prospective studies are needed in the future to confirm and expand these findings.

MULTIPLE EXTRACTION ON PATIENTS WITH DISORDER OF HEMATOPOIESIS (조혈모세포 질환 환자에서의 다발치)

  • Yoon, Hyun-Joong;Kim, Jin
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.27 no.1
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    • pp.87-91
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    • 2001
  • On patients with disorder of hematopoiesis such as leukemia, aplastic anemia, MDS(Myelodisplastic Syndromes), removal of infectious foci prior to the BMT(Bone Marrow Transplantation) is a necessity and what is more, there is no sufficient time to control the infections because the chemotherapy for BMT should be started as soon as possible. And the transfusion should be minimized to prevent the alloimmunization. In those reasons, oral & maxillofacial surgeons are often in need of multiple extractions, and should take into consideration the possibility of complications after multiples extractions such as infection, severe bleeding because those situations can be fatal on patients with disorder of hematopoiesis. We present our experience in multiple extractions on 30 patients with disorder of hematopoiesis referred from Catholic Hematopoiesis Stem Cell transplantation Center at St. Mary's Hospital, The Catholic University of Korea.

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Rapid Micropropagation of Hovenia dulcis Thunb. Through in vitro Stem Nodal Cultures

  • Park, Dong-Jin;Kang, Young-Min;Jung, Ha-Na;Min, Ji-Yun;Kim, Yong-Duck;Karigar, Chandrakant S.;Choi, Myung-Suk
    • Journal of Korean Society of Forest Science
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    • v.95 no.2
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    • pp.155-159
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    • 2006
  • An efficient method for in vitro propagation of the medicinal plant Hovenia duleis, was established. Plantlets for micropropagation of H. dulcis were obtained from in vitro germinated seeds. The effectiveness of various levels of cytokinins (BAP, Kinetin and TDZ) on multiple shoot formation from stem nodes was tested. BAP (1.0 mg/L) treatment induced highest number of multiple shoots. The growth pattern of plantlet on various culture media was undertaken. The shoot elongation was optimal on 2MS basal medium without growth regulators. The in vitro rooting ability of H. dulcis shoots was examined with two-auxins IAA and IBA. The IAA (1.0 mg/L) treatments induced earliest rooting with maximum number of roots and root growth. Rooted shoots were transferred directly to small pots with artificial soil and such established plant exhibited a normal growth pattern similar to wild plantlet.

In vitro Callus formation and Plant Regeneration of Epimedium koreanum Nakai

  • Yu, Chang-Yeon;Kim, Na-Young;Kang, Won-Hee;Kim, Myong-Jo;Heo, Kwon;Choi, Won-Cheol;Lee, Hyeon-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.10 no.4
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    • pp.259-262
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    • 2002
  • The efficiency of regeneration of callus and explants from leaf and stem disks of Epimedium koreanum was examined on the MS media containing 2,4-D, NAA, Kinetin, BA and TDZ. Calli were formed on the 2mg/l 2,4-D media at the rate of 32% from leaf discs and 52% from stems. No callus was produced on the media which are containing BA or TDZ alone. The combination of 2,4-D and BA showed the effect on the formation of callus. The combination of 2mg/l 2,4-D and 0.lmg/l BA in the MS media had produced the highest percentage of callus formation, 50% from leaf discs and 40% from stems, respectively. The combination of 2mg/l 2,4-D and 1mg/1 BA in the MS media had affected the formation of callus in the rate of 40% from leaf discs and 25% from stems. The combined plant growth regulators of 2,4-D and BA increased the formation of calli from leaf discs, but single treatment of 2,4-D showed the highest callus formation from stems. Multiple shoots from leaf discs were formed on the media containing NAA, BA, kinetin, and TDZ. The highest number of multiple shoots were obtained 0.1mg/l NAA combined with 1mg/l kinetin. As a result, leaf discs or stems can be used for the mass propagation of Epimedium koreanum, but stem elongation of shoots from calli was not easy.