Displaying publications 21 - 40 of 216 in total

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  1. Production of bacterial endoglucanase from pretreated oil palm empty fruit bunch by bacillus pumilus EB3
    Ariffin H, Hassan MA, Shah UK, Abdullah N, Ghazali FM, Shirai Y
    J Biosci Bioeng, 2008 Sep;106(3):231-6.
    PMID: 18929997 DOI: 10.1263/jbb.106.231
    In this study, endoglucanase was produced from oil palm empty fruit bunch (OPEFB) by a locally isolated aerobic bacterium, Bacillus pumilus EB3. The effects of the fermentation parameters such as initial pH, temperature, and nitrogen source on the endoglucanase production were studied using carboxymethyl cellulose (CMC) as the carbon source. Endoglucanase from B. pumilus EB3 was maximally secreted at 37 degrees C, initial pH 7.0 with 10 g/l of CMC as carbon source, and 2 g/l of yeast extract as organic nitrogen source. The activity recorded during the fermentation was 0.076 U/ml. The productivity of the enzyme increased twofold when 2 g/l of yeast extract was used as the organic nitrogen supplement as compared to the non-supplemented medium. An interesting finding from this study is that pretreated OPEFB medium showed comparable results to CMC medium in terms of enzyme production with an activity of 0.063 U/ml. As OPEFB is an abundant solid waste at palm oil mills, it has the potential of acting as a substrate in cellulase production.
    Matched MeSH terms: Cell Culture Techniques/methods*
  2. Fulltext Production of Newcastle disease virus by Vero cells grown on cytodex 1 microcarriers in a 2-litre stirred tank bioreactor
    Arifin MA, Mel M, Abdul Karim MI, Ideris A
    J Biomed Biotechnol, 2010;2010:586363.
    PMID: 20625497 DOI: 10.1155/2010/586363
    The aim of this study is to prepare a model for the production of Newcastle disease virus (NDV) lentogenic F strain using cell culture in bioreactor for live attenuated vaccine preparation. In this study, firstly we investigated the growth of Vero cells in several culture media. The maximum cell number was yielded by culture of Vero cells in Dulbecco's Modified Eagle Medium (DMEM) which was 1.93 x 10(6) cells/ml. Secondly Vero cells were grown in two-litre stirred tank bioreactor by using several commercial microcarriers. We achieved the maximum cell concentration about 7.95 x 10(5) cells/ml when using Cytodex 1. Later we produced Newcastle Disease virus in stirred tank bioreactor based on the design developed using Taguchi L4 method. Results reveal that higher multiplicity of infection (MOI) and size of cell inoculums can yield higher virus titer. Finally, virus samples were purified using high-speed centrifugation based on 3( * *)(3-1) Fractional Factorial Design. Statistical analysis showed that the maximum virus titer can be achieved at virus sample concentration of 58.45% (v/v), centrifugation speed of 13729 rpm, and centrifugation time of 4 hours. As a conclusion, high yield of virus titer could be achieved through optimization of cell culture in bioreactor and separation by high-speed centrifugation.
    Matched MeSH terms: Cell Culture Techniques/instrumentation*; Cell Culture Techniques/methods
  3. New method for the isolation of endothelial cells from large vessels
    Ataollahi F, Pingguan-Murphy B, Moradi A, Wan Abas WA, Chua KH, Abu Osman NA
    Cytotherapy, 2014 Aug;16(8):1145-52.
    PMID: 24831838 DOI: 10.1016/j.jcyt.2014.01.010
    Numerous protocols for the isolation of bovine aortic endothelial cells have been described in the previous literature. However, these protocols prevent researchers from obtaining the pure population of endothelial cells. Thus, this study aimed to develop a new and economical method for the isolation of pure endothelial cells by introducing a new strategy to the enzymatic digestion method proposed by previous researchers.
    Matched MeSH terms: Cell Culture Techniques/methods*
  4. Isolation, Expansion, and Characterization of Wharton's Jelly-Derived Mesenchymal Stromal Cell: Method to Identify Functional Passages for Experiments
    Aung SW, Abu Kasim NH, Ramasamy TS
    Methods Mol Biol, 2019;2045:323-335.
    PMID: 31201682 DOI: 10.1007/7651_2019_242
    The therapeutic potential of human mesenchymal stromal stem cells (hMSCs) for cell-based therapeutic is greatly influenced by the in vitro culture condition including the culture conditions. Nevertheless, there are many technical challenges needed to be overcome prior to the clinical use including the quantity, quality, and heterogeneity of the cells. Therefore, it is necessary to develop a stem cell culture procedure or protocol for cell expansion in order to generate reproducible and high-quality cells in accordance with good manufacturing practice for clinical and therapeutic purposes. Here we assessed the MSCs characteristic of human Wharton's jelly mesenchymal stromal cells in in vitro culture according to the criteria established by the International Society for Cellular Therapy. Besides, the viability of the WJMSCs was determined in order to increase the confidence that the cells are employed to meet the therapeutic efficacy.
    Matched MeSH terms: Cell Culture Techniques/methods*
  5. Growth and production of biomass of Rhodovulum sulfidophilum in sardine processing wastewater
    Azad SA, Vikineswary S, Ramachandran KB, Chong VC
    Lett Appl Microbiol, 2001 Oct;33(4):264-8.
    PMID: 11559398
    AIMS: Rhodovulum sulfidophilum was grown in sardine processing wastewater to assess growth characteristics for the production of bacterial biomass with simultaneous reduction of chemical oxygen demand.

    METHODS AND RESULTS: Growth characteristics were compared in diluted and undiluted, settled and non-settled wastewater growing in anaerobic light and aerobic dark conditions; and also at different agitation speeds. The highest biomass (8.75 g l(-1)) and a reduction in chemical oxygen demand of 71% were obtained in unsettled, undiluted wastewater after 120 h culture with 15% inoculum. In settled wastewater, highest biomass (7.64 g l(-1)) and a COD reduction of 77% was also obtained after 120 h. Total biomass was higher (4.34 g l(-1)) after 120 h culture in anaerobic light compared to (3.23 g l(-1)) in aerobic dark growth.

    CONCLUSIONS, SIGNIFICANCE AND IMPACT OF THE STUDY: Better performance, mean of total biomass (6.97 g l(-1) after 96 h), total carotenoids (4.24 mg g(-1) dry cell from 24 h) and soluble protein (431 microg ml(-1) after 96 h) were obtained from aerobic dark culture at 300 rev min(-1). The COD reduction, however, was lower (69%) after 96 h culture. Thus, the benefits in the production of bacterial biomass in non-sterilized sardine processing wastewater with the reduction of chemical oxygen demand could be achieved.

    Matched MeSH terms: Cell Culture Techniques/methods*
  6. Optimization of an induction strategy for improving interferon-alpha2b production in the periplasm of Escherichia coli using response surface methodology
    Azaman SN, Ramakrishnan NR, Tan JS, Rahim RA, Abdullah MP, Ariff AB
    Biotechnol Appl Biochem, 2010 Aug;56(4):141-50.
    PMID: 20604747 DOI: 10.1042/BA20100104
    Induction strategies for the periplasmic production of recombinant human IFN-alpha2b (interferon-alpha2b) by recombinant Escherichia coli Rosetta-gami 2(DE3) were optimized in shake-flask cultures using response surface methodology based on the central composite design. The factors included in the present study were induction point, which related to the attenuance of the cell culture, IPTG (isopropyl beta-D-thiogalactoside) concentration and induction temperature. Second-order polynomial models were used to correlate the abovementioned factors to soluble periplasmic IFN-alpha2b formation and percentage of soluble IFN-alpha2b translocated to the periplasmic space of E. coli. The models were found to be significant and subsequently validated. The proposed induction strategies consisted of induction at an attenuance of 4 (measured as D600), IPTG concentration of 0.05 mM and temperature of 25 degrees C. The optimized induction strategy reduced inclusion-body formation as evidenced by electron microscopy and yielded 323.8 ng/ml of IFN-alpha2b in the periplasmic space with translocation of 74% of the total soluble product. In comparison with the non-optimized condition, soluble periplasmic production and the percentage of soluble IFN-alpha2b translocated to the periplasmic space obtained in optimized induction strategies were increased by approx. 20-fold and 1.4-fold respectively.
    Matched MeSH terms: Cell Culture Techniques/methods*
  7. Prebiotic evaluation of red seaweed (Kappaphycus alvarezii) using in vitro colon model
    Bajury DM, Rawi MH, Sazali IH, Abdullah A, Sarbini SR
    Int J Food Sci Nutr, 2017 Nov;68(7):821-828.
    PMID: 28393631 DOI: 10.1080/09637486.2017.1309522
    Red seaweed (Kappaphycus alvarezii) cultivated from Sabah (RSS) and Langkawi (RSL) were digested using in vitro mouth, gastric and duodenal model. The digested seaweed then fermented in a pH-controlled batch culture system inoculated with human faeces to mimic the distal colon. Bacterial enumeration were monitored using fluorescent in situ hybridisation, and the fermentation end products, the short chain fatty acids (SCFA), were analysed using HPLC. Both RSS and RSL showed significant increase of Bifidobacterium sp.; from log10 7.96 at 0 h to log10 8.72 at 24 h, and from log10 7.96 at 0 h to log10 8.60 at 24 h, respectively, and shows no significant difference when compared to the Bifidobacterium sp. count at 24 h of inulin fermentation. Both seaweeds also showed significant increase in total SCFA production, particularly acetate and propionate. Overall, this data suggested that K. alvarezii might have the potential as a prebiotic ingredient.
    Matched MeSH terms: Batch Cell Culture Techniques
  8. Fulltext Improved Bacteriostatic and Anticorrosion Effects of Polycaprolactone/Chitosan Coated Magnesium via Incorporation of Zinc Oxide
    Bakhsheshi-Rad HR, Hamzah E, Ying WS, Razzaghi M, Sharif S, Ismail AF, et al.
    Materials (Basel), 2021 Apr 12;14(8).
    PMID: 33921460 DOI: 10.3390/ma14081930
    Magnesium has been recognized as a groundbreaking biodegradable biomaterial for implant applications, but its use is limited because it degrades too quickly in physiological solutions. This paper describes the research on the influence of polycaprolactone (PCL)/chitosan (CS)/zinc oxide (ZnO) composite coating (PCL/CS/ZnO) on the corrosion resistance and antibacterial activity of magnesium. The PCL/CS film presented a porous structure with thickness of about 40-50 μm, while after incorporation of ZnO into the PCL/CS, a homogenous film without pores and defects was attained. The ZnO embedded in PCL/CS enhanced corrosion resistance by preventing corrosive ions diffusion in the magnesium substrate. The corrosion, antibacterial, and cell interaction mechanism of the PCL/CS/ZnO composite coating is discussed in this study. In vitro cell culture revealed that the PCL/CS coating with low loaded ZnO significantly improved cytocompatibility, but coatings with high loaded ZnO were able to induce some cytotoxicity osteoblastic cells. It was also found that enhanced antibacterial activity of the PCL/CS/ZnO coating against both Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) bacteria, while less significant antibacterial activity was detected for uncoated Mg and PCL/CS coating. Based on the results, the PCL/CS coatings loaded with low ZnO content may be recommended as a candidate material for biodegradable Mg-based orthopedic implant applications.
    Matched MeSH terms: Cell Culture Techniques
  9. Biologic interaction of three-dimensional periodontal fibroblast spheroids with collagen-based and synthetic membranes
    Berahim Z, Moharamzadeh K, Rawlinson A, Jowett AK
    J. Periodontol., 2011 May;82(5):790-7.
    PMID: 21080786 DOI: 10.1902/jop.2010.100533
    Cell-based therapy using autologous cells has been suggested as a potential approach for periodontal tissue regeneration. Spheroid systems are a form of three-dimensional cell culture that promotes cell matrix interaction, which could recapitulate the aspect of cell homeostasis in vivo. The aim of this study is to assess the interaction of periodontal fibroblast spheroids with synthetic and collagen-based membranes that have been used in guided tissue regeneration.
    Matched MeSH terms: Cell Culture Techniques
  10. A study on the effects of increment and decrement repeated fed-batch feeding of glucose on the production of poly(3-hydroxybutyrate) [P(3HB)] by a newly engineered Cupriavidus necator NSDG-GG mutant in batch fill-and-draw fermentation
    Biglari N, Orita I, Fukui T, Sudesh K
    J Biotechnol, 2020 Jan 10;307:77-86.
    PMID: 31669355 DOI: 10.1016/j.jbiotec.2019.10.013
    This study investigates the effect of strategies on poly(3-hydroxybutyrate) [P(3HB)] production in bioreactor. In the production of P(3HB), urea and glucose feeding streams were developed to characterize the fed-batch culture conditions for new Cupriavidus necator NSDG-GG mutant. Feeding urea in repeated fed-batch stage (RFB-I) at 6, and 12 h in cultivation led to insignificant kinetic effect on the cell dry mass (CDM) and P(3HB) accumulation. Feeding glucose in repeated fed-batch stage (RFB-II) demonstrated that the incremental feeding approach of glucose after urea in fill-and-draw (F/D) mode at 24, 30, 36, 42, and 48 h in fermentation increased CDM and P(3HB) concentration. In the 1st cycle in RFB-II, the cumulative CDM reached the value of 26.22 g/L and then it increased with the successive repeated fed-batches to attain biomass of 145 g/L at the end of 5th cycle of RFB-II. The final cumulative P(3HB) concentration at the end of 5th cycle of RFB-II reached 111 g/L with the overall yield of 0.50 g P(3HB) g gluc- 1; the CDM productivity from the RFB-II cycles was in the range of 0.84-1.3 g/(L·h). The RFB-II of glucose in an increment mode produced nearly 2.2 times more increase in CDM and P(3HB) productivities compared to the decrement RFB-II mode. Repeated cultivation had also the advantage of avoiding extra time required for innoculum preparation, and sterilization of bioreactor during batch, thereby it increased the overall industrial importance of the process.
    Matched MeSH terms: Batch Cell Culture Techniques
  11. Expansion and preservation of multipotentiality of rabbit bone-marrow derived mesenchymal stem cells in dextran-based microcarrier spin culture
    Boo L, Selvaratnam L, Tai CC, Ahmad TS, Kamarul T
    J Mater Sci Mater Med, 2011 May;22(5):1343-56.
    PMID: 21461701 DOI: 10.1007/s10856-011-4294-7
    The use of mesenchymal stem cells (MSCs) in tissue repair and regeneration despite their multipotentiality has been limited by their cell source quantity and decelerating proliferative yield efficiency. A study was thus undertaken to determine the feasibility of using microcarrier beads in spinner flask cultures for MSCs expansion and compared to that of conventional monolayer cultures and static microcarrier cultures. Isolation and characterization of bone marrow derived MSCs were conducted from six adult New Zealand white rabbits. Analysis of cell morphology on microcarriers and culture plates at different time points (D0, D3, D10, D14) during cell culture were performed using scanning electron microscopy and bright field microscopy. Cell proliferation rates and cell number were measured over a period of 14 days, respectively followed by post-expansion characterization. MTT proliferation assay demonstrated a 3.20 fold increase in cell proliferation rates in MSCs cultured on microcarriers in spinner flask as compared to monolayer cultures (p < 0.05). Cell counts at day 14 were higher in those seeded on stirred microcarrier cultures (6.24 ± 0.0420 cells/ml) × 10(5) as compared to monolayer cultures (0.22 ± 0.004 cells/ml) × 10(5) and static microcarrier cultures (0.20 ± 0.002 cells/ml) × 10(5). Scanning electron microscopy demonstrated an increase in cell colonization of the cells on the microcarriers in stirred cultures. Bead-expanded MSCs were successfully differentiated into osteogenic and chondrogenic lineages. This system offers an improved and efficient alternative for culturing MSCs with preservation to their phenotype and multipotentiality.
    Matched MeSH terms: Cell Culture Techniques
  12. Fulltext A Preliminary Study of Human Amniotic Membrane as a Potential Chondrocyte Carrier
    Boo, L., Sofiah, S., Selvaratnam, L., Tai, C.C., Pingguan-Murphy, B., Kamarul, T.
    Malays Orthop J, 2009;3(2):16-23.
    MyJurnal
    Purpose:To investigate the feasibilty of using processed human amniotic membrane (HAM) to support the attachment and proliferation of chondrocytes in vitro which it turn can be utilised as a cell delivery vehicle in tissue engineering applications. Methods: Fresh HAM obtained from patients undergoing routine elective ceasarean sections was harvested., processed and dried using either freez drying (FD) or air drying (AD) methods prior to sterilisation by gamma irradiation. Isolated, processed and characterised rabbit autologous chondrolytes were seeded on processsed HAM and cultured for up to three weeks. Cell attachment and proliferation were examined qualitatively using inverted brightfield microcospy. Results: Processed HAM appeared to allow cell attachment when implanted with chrondocytes. Although cells seeded on AD and FD HAM did not appear to attach as strongly as those seeded on glycerol preserved intact human amniotic membrane, these cells to be proliferated in cell culture conditions. Conclusion: Prelimanary results show that processed HAM chondrocyte attachment and proliferation.
    Matched MeSH terms: Cell Culture Techniques
  13. Fulltext The efficiency of cell sorting and harvesting methods for In vitro expansion of human umbilical cord blood derived CD34+ hematopoietic stem cells
    Borojerdi, Mohadese Hashem, Maqbool, Maryam, Zuraidah Yusoff, Vidyadaran, Sharmili, Hwa, Ling King, George, Elizabeth, et al.
    Malaysian Journal of Medicine and Health Sciences, 2015;11(2):21-28.
    MyJurnal
    Introduction: During the last three decades hematopoietic stem cell transplantation (HSCT) has become a well-established treatment for many hematologic malignancies. The most important limitation for HSC transplantation is the low number of hematopoietic stem cells (HSC) that can lead to delayed engraftment or graft failures. Numerous attempts have been made to improve in vitro HSC expansion via optimization of various methods such as isolation techniques, supplementing with growth factors, utilizing stromal cells as feeder layer and other culture conditions. Objective: This project is aimed to decipher the efficiency of an isolation technique and retrieval of culture expanded HSC from feeder layer using two different harvesting methods. Materials and Methods: Hematopoietic stem cells from human umbilical cord blood were isolated via MACS mediated CD34+ double sorting. Then, the cells were cultured onto MSC feeder layer for 3 and 5 days. Culture expanded cells were harvested using two different harvesting method namely cell aspiration and trypsinization methods. Hematopoietic stem cell expansion index were calculated based on harvesting methods for each time point. Results: The numbers of HSC isolated from human umbilical cord blood were 1.64 x 106 and 1.20 x106 cells at single and double sortings respectively. Although the number of sorted cells diminished at the second sorting yet the yield of CD34+ purity has increased from 43.73% at single sorting to 81.40% at double sorting. Employing the trypsinization method, the HSC harvested from feeder layer showed a significant increase in expansion index (EI) as compared to the cell aspiration harvesting method (p≤ 0.05). However, the purity of CD34+ HSC was found higher when the cells were harvested using aspiration method (82.43%) as compared to the trypsinization method (74.13%). Conclusion: A pure population of CD34+ HSC can be retrieved when the cells were double sorted using MACS and expanded in culture after being harvested using cell aspiration method.
    Matched MeSH terms: Cell Culture Techniques
  14. Tissue-Engineered Skin Substitute Enhances Wound Healing after Radiation Therapy
    Busra MF, Chowdhury SR, bin Ismail F, bin Saim A, Idrus RB
    Adv Skin Wound Care, 2016 Mar;29(3):120-9.
    PMID: 26866868 DOI: 10.1097/01.ASW.0000480556.78111.e4
    OBJECTIVE: When given in conjunction with surgery for treating cancer, radiation therapy may result in impaired wound healing, which, in turn, could cause skin ulcers. In this study, bilayer and monolayer autologous skin substitutes were used to treat an irradiated wound.

    MATERIALS AND METHODS: A single dose of 30 Gy of linear electron beam radiation was applied to the hind limb of nude mice before creating the skin lesion (area of 78.6 mm). Monolayer tissue-engineered skin substitutes (MTESSs) were prepared by entrapping cultured keratinocytes in fibrin matrix, and bilayer tissue-engineered skin substitutes (BTESSs) were prepared by entrapping keratinocytes and fibroblasts in separate layers. Bilayer tissue-engineered skin substitute and MTESS were implanted to the wound area. Gross appearance and wound area were analyzed to evaluate wound healing efficiency. Skin regeneration and morphological appearance were observed via histological and electron microscopy. Protein expressions of transforming growth factor β1 (TGF-β1), platelet-derived growth factor BB (PDGF-BB), and vascular endothelial growth factor (VEGF) in skin regeneration were evaluated by immunohistochemistry (IHC).

    RESULTS: Macroscopic observation revealed that at day 13, treatments with BTESS completely healed the irradiated wound, whereas wound sizes of 1.1 ± 0.05 and 6.8 ± 0.14 mm were measured in the MTESS-treated and untreated control groups, respectively. Hematoxylin-eosin (H&E) analysis showed formation of compact and organized epidermal and dermal layers in the BTESS-treated group, as compared with MTESS-treated and untreated control groups. Ultrastructural analysis indicates maturation of skin in BTESS-treated wound evidenced by formation of intermediate filament bundles in the dermal layer and low intercellular space in the epidermal layer. Expressions of TGF-β1, PDGF-BB, and VEGF were also higher in BTESS-treated wounds, compared with MTESS-treated wounds.

    CONCLUSIONS: These results indicate that BTESS is the preferred treatment for irradiated wound ulcers.

    Matched MeSH terms: Cell Culture Techniques
  15. Development of a novel model for the investigation of implant-soft tissue interface
    Chai WL, Moharamzadeh K, Brook IM, Emanuelsson L, Palmquist A, van Noort R
    J. Periodontol., 2010 Aug;81(8):1187-95.
    PMID: 20450401 DOI: 10.1902/jop.2010.090648
    In dental implant treatment, the long-term prognosis is dependent on the biologic seal formed by the soft tissue around the implant. The in vitro investigation of the implant-soft tissue interface is usually carried out using a monolayer cell-culture model that lacks a polarized-cell phenotype. This study developed a tissue-engineered three-dimensional oral mucosal model (3D OMM) to investigate the implant-soft tissue interface.
    Matched MeSH terms: Cell Culture Techniques
  16. Fulltext Effects of abiotic stress on biomass and anthocyanin production in cell cultures of Melastoma malabathricum
    Chan LK, Koay SS, Boey PL, Bhatt A
    Biol Res, 2010;43(1):127-35.
    PMID: 21157639 DOI: /S0716-97602010000100014
    Plant cell cultures could be used as an important tool for biochemical production, ranging from natural coloring (pigments) to pharmaceutical products. Anthocyanins are becoming a very important alternative to synthetic dyes because of increased public concern over the safety of artificial food coloring agents. Several factors are responsible for the production of anthocyanin in cell cultures. In the present study, we investigate the effects of different environmental factors, such as light intensity, irradiance (continuous irradiance or continuous darkness), temperature and medium pH on cell biomass yield and anthocyanin production in cultures of Melastoma malabathricum. Moderate light intensity (301 - 600 lux) induced higher accumulation of anthocyanins in the cells. The cultures exposed to 10-d continuous darkness showed the lowest pigment content, while the cultures exposed to 10-d continuous irradiance showed the highest pigment content. The cell cultures incubated at a lower temperature range (20 ± 2 ºC) grew better and had higher pigment content than those grown at 26 ± 2 ºC and 29 ± 2 ºC. Different medium pH did not affect the yield of cell biomass but anthocyanin accumulation was highest at pH 5.25 - 6.25.
    Matched MeSH terms: Cell Culture Techniques
  17. Larvicidal, oviposition, and ovicidal effects of Artemisia annua (Asterales: Asteraceae) against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus (Diptera: Culicidae)
    Cheah SX, Tay JW, Chan LK, Jaal Z
    Parasitol Res, 2013 Sep;112(9):3275-82.
    PMID: 23835922 DOI: 10.1007/s00436-013-3506-0
    This study focuses on the larvicidal, oviposition, and ovicidal effects of a crude extract of Artemisia annua against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus. Dried cells of Artemisia annua from cell suspension cultures were extracted using hexane. The extract showed moderate larvicidal effects against mosquitoes. At 24-h post treatment, the LC50 values for Anopheles sinensis, Aedes aegypti, and Culex quinquefasciatus were recorded as 244.55, 276.14, and 374.99 ppm, respectively. The percentage mortality of larvae was directly proportional to the tested concentration. Anopheles sinensis was found to be the most susceptible species, whereas Culex quinquefasciatus was the most tolerant to the Artemisia annua extract. The results indicated that the Artemisia annua extract showed concentration-dependent oviposition deterrent activity and had a strong deterrent effect. At 500 ppm, the percentage effective repellency was more than 85% compared with the control group for all the species, with oviposition activity index values of -0.94, -0.95, and -0.78 for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. In the ovicidal assay, the percentage hatchability of eggs after treatment with 500 ppm of Artemisia annua extract was significantly lower than the control, with values of 48.84 ± 4.08, 38.42 ± 3.67, and 79.35 ± 2.09% for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. Artemisia annua was found to be more effective against Aedes aegypti and Anopheles sinensis compared with Culex quinquefasciatus. This study indicated that crude extract of A. annua could be a potential alternative for use in vector management programs.
    Matched MeSH terms: Cell Culture Techniques
  18. Xeno-free and feeder-free culture and differentiation of human embryonic stem cells on recombinant vitronectin-grafted hydrogels
    Chen LH, Sung TC, Lee HH, Higuchi A, Su HC, Lin KJ, et al.
    Biomater Sci, 2019 Aug 14.
    PMID: 31411209 DOI: 10.1039/c9bm00418a
    Recombinant vitronectin-grafted hydrogels were developed by adjusting surface charge of the hydrogels with grafting of poly-l-lysine for optimal culture of human embryonic stem cells (hESCs) under xeno- and feeder-free culture conditions, with elasticity regulated by crosslinking time (10-30 kPa), in contrast to conventional recombinant vitronectin coating dishes, which have a fixed stiff surface (3 GPa). hESCs proliferated on the hydrogels for over 10 passages and differentiated into the cells derived from three germ layers indicating the maintenance of pluripotency. hESCs on the hydrogels differentiated into cardiomyocytes under xeno-free culture conditions with much higher efficiency (80% of cTnT+ cells) than those on conventional recombinant vitronectin or Matrigel-coating dishes just only after 12 days of induction. It is important to have an optimal design of cell culture biomaterials where biological cues (recombinant vitronectin) and physical cues (optimal elasticity) are combined for high differentiation of hESCs into specific cell lineages, such as cardiomyocytes, under xeno-free and feeder-free culture conditions.
    Matched MeSH terms: Cell Culture Techniques
  19. Exploring the additive bio-agent impacts upon ectoine production by Halomonas salina DSM5928T using corn steep liquor and soybean hydrolysate as nutrient supplement
    Chen PW, Cui ZY, Ng HS, Chi-Wei Lan J
    J Biosci Bioeng, 2020 Aug;130(2):195-199.
    PMID: 32370929 DOI: 10.1016/j.jbiosc.2020.03.011
    Ectoine production using inexpensive and renewable biomass resources has attracted great interest among the researchers due to the low yields of ectoine in current fermentation approaches that complicate the large-scale production of ectoine. In this study, ectoine was produced from corn steep liquor (CSL) and soybean hydrolysate (SH) in replacement to yeast extract as the nitrogen sources for the fermentation process. To enhance the bacterial growth and ectoine production, biotin was added to the Halomonas salina fermentation media. In addition, the effects addition of surfactants such as Tween 80 and saponin on the ectoine production were also investigated. Results showed that both the CSL and SH can be used as the nitrogen source substitutes in the fermentation media. Higher amount of ectoine (1781.9 mg L-1) was produced in shake flask culture with SH-containing media as compared to CSL-containing media. A total of 2537.0 mg L-1 of ectoine was produced at pH 7 when SH-containing media was applied in the 2 L batch fermentation. Moreover, highest amount of ectoine (1802.0 mg L-1) was recorded in the SH-containing shake flask culture with addition of 0.2 μm mL-1 biotin. This study demonstrated the efficacy of industrial waste as the nutrient supplement for the fermentation of ectoine production.
    Matched MeSH terms: Batch Cell Culture Techniques
  20. Statistical optimization of green fluorescent protein production from Escherichia coli BL21(DE3)
    Chew FN, Tan WS, Boo HC, Tey BT
    Prep Biochem Biotechnol, 2012;42(6):535-50.
    PMID: 23030465 DOI: 10.1080/10826068.2012.660903
    An optimized cultivation condition is needed to maximize the functional green fluorescent protein (GFP) production. Six process variables (agitation rate, temperature, initial medium pH, concentration of inducer, time of induction, and inoculum density) were screened using the fractional factorial design. Three variables (agitation rate, temperature, and time of induction) exerted significant effects on functional GFP production in E. coli shake flask cultivation and were optimized subsequently using the Box-Behnken design. An agitation rate of 206 rpm at 31°C and induction of the protein expression when the cell density (OD(600nm)) reaches 1.04 could enhance the yield of functional GFP production from 0.025 g/L to 0.241 g/L, which is about ninefold higher than the unoptimized conditions. Unoptimized cultivation conditions resulted in protein aggregation and hence reduced the quantity of functional GFP. The model and regression equation based on the shake flask cultivation could be applied to a 2-L bioreactor for maximum functional GFP production.
    Matched MeSH terms: Cell Culture Techniques/methods; Cell Culture Techniques/standards
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