Displaying publications 161 - 180 of 315 in total

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  1. Fulltext Long-term xeno-free culture of human pluripotent stem cells on hydrogels with optimal elasticity
    Higuchi A, Kao SH, Ling QD, Chen YM, Li HF, Alarfaj AA, et al.
    Sci Rep, 2015 Dec 14;5:18136.
    PMID: 26656754 DOI: 10.1038/srep18136
    The tentative clinical application of human pluripotent stem cells (hPSCs), such as human embryonic stem cells and human induced pluripotent stem cells, is restricted by the possibility of xenogenic contamination resulting from the use of mouse embryonic fibroblasts (MEFs) as a feeder layer. Therefore, we investigated hPSC cultures on biomaterials with different elasticities that were grafted with different nanosegments. We prepared dishes coated with polyvinylalcohol-co-itaconic acid hydrogels grafted with an oligopeptide derived from vitronectin (KGGPQVTRGDVFTMP) with elasticities ranging from 10.3 to 30.4 kPa storage moduli by controlling the crosslinking time. The hPSCs cultured on the stiffest substrates (30.4 kPa) tended to differentiate after five days of culture, whereas the hPSCs cultured on the optimal elastic substrates (25 kPa) maintained their pluripotency for over 20 passages under xeno-free conditions. These results indicate that cell culture matrices with optimal elasticity can maintain the pluripotency of hPSCs in culture.
    Matched MeSH terms: Cell Culture Techniques/methods*
  2. Fulltext Laminin-Coated Poly(Methyl Methacrylate) (PMMA) Nanofiber Scaffold Facilitates the Enrichment of Skeletal Muscle Myoblast Population
    Zahari NK, Idrus RBH, Chowdhury SR
    Int J Mol Sci, 2017 Oct 30;18(11).
    PMID: 29084180 DOI: 10.3390/ijms18112242
    Myoblasts, the contractile cells of skeletal muscle, have been invaluable for fundamental studies of muscle development and clinical applications for muscle loss. A major limitation to the myoblast-based therapeutic approach is contamination with non-contractile fibroblasts, which overgrow during cell expansion. To overcome these limitations, this study was carried out to establish a 3D culture environment using nanofiber scaffolds to enrich the myoblast population during construct formation. Poly(methyl methacrylate) (PMMA) nanofiber (PM) scaffolds were fabricated using electrospinning techniques and coated with extracellular matrix (ECM) proteins, such as collagen or laminin, in the presence or absence of genipin. A mixed population of myoblasts and fibroblasts was isolated from human skeletal muscle tissues and cultured on plain surfaces, as well as coated and non-coated PM scaffolds. PMMA can produce smooth fibers with an average diameter of 360 ± 50 nm. Adsorption of collagen and laminin on PM scaffolds is significantly enhanced in the presence of genipin, which introduces roughness to the nanofiber surface without affecting fiber diameter and mechanical properties. It was also demonstrated that laminin-coated PM scaffolds significantly enhance myoblast proliferation (0.0081 ± 0.0007 h-1) and migration (0.26 ± 0.04 μm/min), while collagen-coated PM scaffolds favors fibroblasts proliferation (0.0097 ± 0.0009 h-1) and migration (0.23 ± 0.03 μm/min). Consequently, the myoblast population was enriched on laminin-coated PM scaffolds throughout the culture process. Therefore, laminin coating of nanofiber scaffolds could be a potential scaffold for the development of a tissue-engineered muscle substitute.
    Matched MeSH terms: Cell Culture Techniques
  3. Fulltext Prevalence and Molecular Aspects of Human Hookworms in Guilan Province, Northern Iran
    Sharifdini M, Ghanbarzadeh L, Kouhestani-Maklavani N, Mirjalali H, Saraei M
    Iran J Parasitol, 2017 Jul-Sep;12(3):374-381.
    PMID: 28979347
    BACKGROUND: Hookworm infection is one of the important Neglected Tropical Diseases (NTD) in the world. It was previously more prevalent in the northern and southern parts of Iran with a prevalence rate higher than 40% in some endemic regions; nevertheless, the infection rate has decreased to less than 1%. This study aimed to determine prevalence and molecular aspects of hookworm infections in rural inhabitants of Fouman County, Guilan Province, northern Iran.

    METHODS: This cross-sectional study was performed in 31 villages of Fouman district in Guilan Province, northern Iran during 2015-2016. Stool samples were collected from 1500 rural inhabitants and examined by formalin ethyl-acetate concentration as well as agar plate culture techniques. After treatment with albendazole, adult hookworms were isolated. Following DNA extraction, PCR amplification of ITS2-rDNA region was performed and the product was sequenced, followed by genetic variation analysis.

    RESULTS: Of 1500 samples, one case was morphologically diagnosed as N. americanus. In addition, molecular characterization verified the presence of N. americanus, showing more than 95% similarity with sequences of N. americanus present in GenBank. The patient showed no clinical symptoms and a mild hypereosinophilia was the only laboratory finding observed.

    CONCLUSION: A reduced prevalence of human hookworms was demonstrated within Guilan Province located in north of Iran. The N. americanus originated from Guilan had a high homology with the isolates found in Japan, Laos, Malaysia, and Australia.
    Matched MeSH terms: Culture Techniques
  4. Pooled Human Serum Increases Regenerative Potential of In Vitro Expanded Stem Cells from Human Extracted Deciduous Teeth
    Haque N, Abu Kasim NH
    Adv Exp Med Biol, 2017 7 22;1083:29-44.
    PMID: 28730381 DOI: 10.1007/5584_2017_74
    In regenerative therapy, in vitro expansion of stem cells is critical to obtain a significantly higher number of cells for successful engraftment after transplantation. However, stem cells lose its regenerative potential and enter senescence during in vitro expansion. In this study, the influence of foetal bovine serum (FBS) and pooled human serum (pHS) on the proliferation, morphology and migration of stem cells from human extracted deciduous teeth (SHED) was compared. SHED (n = 3) was expanded in KnockOut DMEM supplemented with either pHS (pHS-SM) or FBS (FBS-SM). pHS was prepared using peripheral blood serum of six healthy male adults, aged between 21 and 35 years old. The number of live SHED was significantly higher, from passage 5 to 7, when cultured in pHS-SM compared to those cultured in FBS-SM (p 
    Matched MeSH terms: Cell Culture Techniques
  5. Fulltext Macrophage-derived interleukin-1beta promotes human breast cancer cell migration and lymphatic adhesion in vitro
    Storr SJ, Safuan S, Ahmad N, El-Refaee M, Jackson AM, Martin SG
    Cancer Immunol Immunother, 2017 Oct;66(10):1287-1294.
    PMID: 28551814 DOI: 10.1007/s00262-017-2020-0
    Lymphovascular invasion (LVI), encompassing blood and lymphatic vessel invasion, is an important event in tumourigenesis. Macrophages within the tumour microenvironment are linked to the presence of LVI and angiogenesis. This study investigates the role of macrophage-derived, caspase-1-dependent interleukin-1beta (IL-1β) in an in vitro model of LVI. IL-1β significantly augmented the adhesion and transmigration of breast cancer cell lines MCF7 and MDA-MB-231 across endothelial cell barriers. MDA-MB-231 and MCF7 showed a higher percentage of adhesion to lymphatic endothelial cells than blood endothelial cells following endothelial cell IL-1β stimulation (P 
    Matched MeSH terms: Cell Culture Techniques
  6. An evaluation of crude palm oil (CPO) and tocotrienol rich fraction (TRF) of palm oil as percutaneous permeation enhancers using full-thickness human skin
    Singh I, Nair RS, Gan S, Cheong V, Morris A
    Pharm Dev Technol, 2019 Apr;24(4):448-454.
    PMID: 30084268 DOI: 10.1080/10837450.2018.1509347
    The drawbacks associated with chemical skin permeation enhancers such as skin irritation and toxicity necessitated the research to focus on potential permeation enhancers with a perceived lower toxicity. Crude palm oil (CPO) is obtained by direct compression of the mesocarp of the fruit of the oil palm belonging to the genus Elaeis. In this research, CPO and tocotrienol-rich fraction (TRF) of palm oil were evaluated for the first time as skin permeation enhancers using full-thickness human skin. The in vitro permeation experiments were conducted using excised human skin mounted in static upright 'Franz-type' diffusion cells. The drugs selected to evaluate the enhancing effects of these palm oil derivatives were 5-fluorouracil, lidocaine and ibuprofen: compounds covering a wide range of Log p values. It was demonstrated that CPO and TRF were capable of enhancing the percutaneous permeation of drugs across full-thickness human skin in vitro. Both TRF and CPO were shown to significantly enhance the permeation of ibuprofen with flux values of 30.6 µg/cm2 h and 23.0 µg/cm2 h respectively, compared to the control with a flux of 16.2 µg/cm2 h. The outcome of this research opens further scope for investigation on the transdermal penetration enhancement activity of pure compounds derived from palm oil.
    Matched MeSH terms: Organ Culture Techniques
  7. Fluorescent cell tracer dye permits real-time assessment of re-epithelialization in a serum-free ex vivo human skin wound assay
    Nasir NAM, Paus R, Ansell DM
    Wound Repair Regen, 2019 01;27(1):126-133.
    PMID: 30575205 DOI: 10.1111/wrr.12688
    Ex vivo wounded human skin organ culture is an invaluable tool for translationally relevant preclinical wound healing research. However, studies incorporating this system are still underutilized within the field because of the low throughput of histological analysis required for downstream assessment. In this study, we use intravital fluorescent dye to lineage trace epidermal cells, demonstrating that wound re-epithelialization of human ex vivo wounds occurs consistent with an extending shield mechanism of collective migration. Moreover, we also report a relatively simple method to investigate global epithelial closure of explants in culture using daily fluorescent dye treatment and en face imaging. This study is the first to quantify healing of ex vivo wounds in a longitudinal manner, providing global assessments for re-epithelialization and tissue contraction. We show that this approach can identify alterations to healing with a known healing promoter. This methodological study highlights the utility of human ex vivo wounds in enhancing our understanding of mechanisms of human skin repair and in evaluating novel therapies to improve healing outcome.
    Matched MeSH terms: Organ Culture Techniques
  8. Establishment of epithelial and fibroblast cell cultures and cell lines from primary renal cancer nephrectomies
    Yap NY, Ong TA, Morais C, Pailoor J, Gobe GC, Rajandram R
    Cell Biol Int, 2019 Jun;43(6):715-725.
    PMID: 31062478 DOI: 10.1002/cbin.11150
    Renal cell carcinoma (RCC) is one of the most lethal urogenital cancers and effective treatment of metastatic RCC remains an elusive target. Cell lines enable the in vitro investigation of molecular and genetic changes leading to renal carcinogenesis and are important for evaluating cellular drug response or toxicity. This study details a fast and easy protocol of establishing epithelial and fibroblast cell cultures or cell lines concurrently from renal cancer nephrectomy tissue. The protocol involves mechanical disaggregation, collagenase digestion and cell sieving for establishing epithelial cells while fibroblast cells were grown from explants. This protocol has been modified from previous published reports with additional antibiotics and washing steps added to eliminate microbial contamination from the surgical source. Cell characterisation was carried out using immunofluorescence and quantitative polymerase chain reaction. Eleven stable epithelial renal tumour cell lines of various subtypes, including rare subtypes, were established with a spontaneous immortalisation rate of 21.6% using this protocol. Eight fibroblast cell cultures grew successfully but did not achieve spontaneous immortalisation. Cells of epithelial origin expressed higher expressions of epithelial markers such as pan-cytokeratin, cytokeratin 8 and E-cadherin whereas fibroblast cells expressed high α-smooth muscle actin. Further mutational analysis is needed to evaluate the genetic or molecular characteristics of the cell lines.
    Matched MeSH terms: Cell Culture Techniques
  9. 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
  10. 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
  11. Fulltext Enhancement of β-Mannanase Production by Bacillus subtilis ATCC11774 through Optimization of Medium Composition
    Norizan NABM, Halim M, Tan JS, Abbasiliasi S, Mat Sahri M, Othman F, et al.
    Molecules, 2020 Jul 31;25(15).
    PMID: 32752106 DOI: 10.3390/molecules25153516
    Palm kernel cake (PKC) has been largely produced in Malaysia as one of the cheap and abundant agro-waste by-products from the palm oil industry and it contains high fiber (mannan) content. The present study aimed to produce β-mannanase by Bacillus subtilis ATCC11774 via optimization of the medium composition using palm kernel cake as substrate in semi-solid fermentation. The fermentation nutrients such as PKC, peptone, yeast extract, sodium chloride, magnesium sulphate (MgSO2), initial culture pH and temperature were screened using a Plackett-Burman design. The three most significant factors identified, PKC, peptone and NaCl, were further optimized using central composite design (CCD), a response surface methodology (RSM) approach, where yeast extract and MgSO2 were fixed as a constant factor. The maximum β-mannanase activity predicted by CCD under the optimum medium composition of 16.50 g/L PKC, 19.59 g/L peptone, 3.00 g/L yeast extract, 2.72 g/L NaCl and 0.2 g/L MgSO2 was 799 U/mL. The validated β-mannanase activity was 805.12 U/mL, which was close to the predicted β-mannanas activity. As a comparison, commercial media such as nutrient broth, M9 and Luria bertani were used for the production of β-mannanase with activities achieved at 204.16 ± 9.21 U/mL, 50.32 U/mL and 88.90 U/mL, respectively. The optimized PKC fermentation medium was four times higher than nutrient broth. Hence, it could be a potential fermentation substrate for the production of β-mannanase activity by Bacillus subtilis ATCC11774.
    Matched MeSH terms: Batch Cell Culture Techniques
  12. Fulltext Detection of dengue virus from field Aedes aegypti and Aedes albopictus adults and larvae
    Ahmad R, Ismail A, Saat Z, Lim LH
    Southeast Asian J Trop Med Public Health, 1997 Mar;28(1):138-42.
    PMID: 9322296
    Mosquito adults and larvae were collected from dengue high risk areas and transported to the laboratory for identification. Identified mosquitos were pooled according to the species, date and locality and stored at -70 degrees C. A total of 1,385 pools of Aedes albopictus and 267 pools of Ae. Aegypti were collected from major towns in 12 states in Peninsular Malaysia. Virus isolation was carried out using cell culture (C6/36 clone) of Ae. albopictus and detection of dengue virus by the peroxidase anti-peroxidase staining. All positive isolations were further re-confirmed by the reverse transcriptase-polymerase chain reaction (RT-PCR). Most of the pools were negative with PAP staining and RT-PCR. However, 11 mosquito pools were positive with PAP staining. On the other hand, samples from Terengganu, Pulau Pinang and Johor were positive using both methods.
    Matched MeSH terms: Cell Culture Techniques
  13. Evaluation of different promoters driving the GFP reporter gene and selected target tissues for particle bombardment of Dendrobium Sonia 17
    Tee CS, Marziah M, Tan CS, Abdullah MP
    Plant Cell Rep, 2003 Jan;21(5):452-8.
    PMID: 12789448
    Three different morphological callus types, identified as type A, B and C, and tips of in vitro inflorescences were used as target tissues for genetic transformation. Five different DNA plasmids carrying a synthetic green fluorescent protein (gfp) gene driven by different promoters, CaMV 35S, HBT, and Ubi1 were tested for the genetic transformation of Dendrobium Sonia 17. 35S-sgfp-TYG-nos (p35S) with the CaMV 35S promoter showed the highest GFP transient expression rate, while the HBT and Ubi1 promoters showed a relatively lower expression rate in all of the target tissues tested. The highest number of GFP-expressing cells was observed on day 2 post-bombardment, and the number declined gradually over the course of the next 2 weeks. Type A and B callus were found to be the best potential target tissues for genetic transformation.
    Matched MeSH terms: Culture Techniques
  14. Strategy for generating tissue-engineered human bone construct
    Tan KK, Aminuddin BS, Tan GH, Sabarul Afian M, Ng MH, Fauziah O, et al.
    Med J Malaysia, 2004 May;59 Suppl B:43-4.
    PMID: 15468810
    The strategy used to generate tissue-engineered bone construct, in view of future clinical application is presented here. Osteoprogenitor cells from periosteum of consenting scoliosis patients were isolated. Growth factors viz TGF-B2, bFGF and IGF-1 were used in concert to increase cell proliferation during in vitro cell expansion. Porous tricalcium phosphate (TCP)-hydroxyapatite (HA) scaffold was used as the scaffold to form 3D bone construct. We found that the addition of growth factors, greatly increased cell growth by 2 to 7 fold. TCP/HA proved to be the ideal scaffold for cell attachment and proliferation. Hence, this model will be further carried out on animal trial.
    Matched MeSH terms: Organ Culture Techniques
  15. Effects of Melatonin on Colchicine-Treated PLBs of Dendrobium sonia-28 Orchid
    Lim MS, Antony JJ, Islam SM, Suhana Z, Sreeramanan S
    Appl Biochem Biotechnol, 2017 Jan;181(1):15-31.
    PMID: 27461541 DOI: 10.1007/s12010-016-2196-3
    Dendrobium hybrid orchid is popular in orchid commercial industry due to its short life cycle and ability to produce various types of flower colours. This study was conducted to identify the morphological, biochemical and scanning electron microscopy (SEM) analysis in the Dendrobium sonia-28 orchid plants. In this study, 0.05 and 0.075 % of colchicine-treated Dendrobium sonia-28 (4-week-old culture) protocorm-like bodies (PLBs) were treated in different concentrations of melatonin (MEL) posttreatments (0, 0.05, 0.1, 0.5, 1, 5 and 10 μM). Morphological parameters such as number of shoots, growth index and number of PLBs were determined. In the 0.05 and 0.075 % of colchicine-treated PLBs which were posttreated with 0.05 μM MEL resulted in the highest value of the morphological parameters tested based on the number of shoots (84.5 and 96.67), growth index (16.94 and 12.15) and number of PLBs (126.5 and 162.33), respectively. SEM analysis of the 0.05 μM MEL posttreatment on both the colchicine-treated regenerated PLBs showed irregular cell lineages, and some damages occurred on the stomata. This condition might be due to the effect of plasmolyzing occurred in the cell causing irregular cell lineages.
    Matched MeSH terms: Cell Culture Techniques
  16. Triatomines as experimental vectors of trypanosomes of Asian monkeys
    Weinman D, Wallis RC, Cheong WH, Mahadevan S
    Am J Trop Med Hyg, 1978 Mar;27(2 Pt 1):232-7.
    PMID: 417639
    Systematic surveys of the wild macaques of South Asia by blood culture resulted in the discovery that trypanosomiasis is enzootic in the simians of Indonesia, Malaysia, India, and Thailand. The isolates obtained differ in morphology, metabolism, and ability to multiply in arthropods. Following this discovery, interest focused on possible transmissions of these trypanosomiases. Laboratory-reared and wild-caught insects were studied to determine which are satisfactory intermediate hosts and potential natural vectors. Successful results were obtained with insectary-reared reduviids and Indonesian isolates. In Rhodnius prolixus and Triatoma rubrofasciata the Indonesian trypanosomes multiply for periods which can exceed 40 days. The flagellate infections are in the digestive tract, whereas trypanosomes have never been seen in the salivary glands or in the hemolymph. The feces of trypanosome-carrying reduviids are infective, suggesting a stercoreal method of infection of mammals, and infection was produced in experiments in which feeding by the insects was not possible. The relevance of these findings to natural transmission in Indonesia is not known. Experiments with insects and all other trypanosomal isolates have been negative. The natural transmission mechanism(s) of the simian trypanosomiases in South Asia remains an unsolved problem.
    Matched MeSH terms: Culture Techniques
  17. Biomechanical comparison between cortical screw-rod construct versus pedicle screw-rod construct in transforaminal lumbar interbody fusion
    Yin Wei CC, Haw SS, Bashir ES, Beng SL, Shanmugam R, Keong KM
    J Orthop Surg (Hong Kong), 2017 01;25(1):2309499017690656.
    PMID: 28219305 DOI: 10.1177/2309499017690656
    OBJECTIVE: To compare construct stiffness of cortical screw (CS)-rod transforaminal lumbar interbody fusion (TLIF) construct (G2) versus pedicle screw (PS)-rod TLIF construct (G1) in the standardized porcine lumbar spine.

    METHODS: Six porcine lumbar spines (L2-L5) were separated into 12 functional spine units. Bilateral total facetectomies and interlaminar decompression were performed for all specimens. Non-destructive loading to assess stiffness in lateral bending, flexion and extension as well as axial rotation was performed using a universal material testing machine.

    RESULTS: PS and CS constructs were significantly stiffer than the intact spine except in axial rotation. Using the normalized ratio to the intact spine, there is no significant difference between the stiffness of PS and CS: flexion (1.41 ± 0.27, 1.55 ± 0.32), extension (1.98 ± 0.49, 2.25 ± 0.44), right lateral flexion (1.93 ± 0.57, 1.55 ± 0.30), left lateral flexion (2.00 ± 0.73, 2.16 ± 0.20), right axial rotation (0.99 ± 0.21, 0.83 ± 0.26) and left axial rotation (0.96 ± 0.22, 0.92 ± 0.25).

    CONCLUSION: The CS-rod TLIF construct provided comparable construct stiffness to a traditional PS-rod TLIF construct in a 'standardized' porcine lumbar spine model.

    Matched MeSH terms: Tissue Culture Techniques
  18. Fulltext Establishing a culture system that supports in vitro expansion of adult microglia
    Subramaniam, Hemavathy, Alireza Badiei, Ramasamy, Rajesh, Maha Abdullah, Vidyadaran, Sharmili
    Malaysian Journal of Medicine and Health Sciences, 2010;6(2):25-30.
    MyJurnal
    Introduction: The vast majority of in vitro research on microglia are based on cells isolated from
    neonatal animals (3-5 days of age). Studying microglia of adults has been limited by the lack of a suitable culture system that supports their growth. In this study, we describe a protocol for growing microglia of adults based on modifications of the technique for culturing microglia isolated from neonatal rats. Methods: Mixed glia isolated from adult rats (age range of 1 month to 3 years old) were seeded in
    culture flasks coated with poly-L-lysine. Cells were maintained in DMEM media supplemented with
    insulin-transferrin-selenium (ITS) and recombinant human macrophage colony-stimulating factor
    (M-CSF). Mild trypsinisation was carried out to isolate microglia from mixed glia culture. Results:
    Microglia cells of adult rats were successfully grown in vitro. For the expansion of adult microglia,
    it was observed that coating the cell culture flasks with poly-L-lysine was crucial to encourage cell
    adherence. The substitution of insulin in culture media with ITS was found to improve cell yield and
    reduced the number of days required for culture from 28 days to 14 days. Addition of M-CSF to cell
    culture medium, along with the improvisations described above provided the best adult microglia cell
    yield (2.91 ± 0.56 x 106 cells) compared to the technique of replating cells (0.91 ± 0.65 x 106 cells;
    p
    Matched MeSH terms: Cell Culture Techniques
  19. Tissue-engineered trachea: A review
    Law JX, Liau LL, Aminuddin BS, Ruszymah BH
    Int J Pediatr Otorhinolaryngol, 2016 Dec;91:55-63.
    PMID: 27863642 DOI: 10.1016/j.ijporl.2016.10.012
    Tracheal replacement is performed after resection of a portion of the trachea that was impossible to reconnect via direct anastomosis. A tissue-engineered trachea is one of the available options that offer many advantages compared to other types of graft. Fabrication of a functional tissue-engineered trachea for grafting is very challenging, as it is a complex organ with important components, including cartilage, epithelium and vasculature. A number of studies have been reported on the preparation of a graftable trachea. A laterally rigid but longitudinally flexible hollow cylindrical scaffold which supports cartilage and epithelial tissue formation is the key element. The scaffold can be prepared via decellularization of an allograft or fabricated using biodegradable or non-biodegradable biomaterials. Commonly, the scaffold is seeded with chondrocytes and epithelial cells at the outer and luminal surfaces, respectively, to hasten tissue formation and improve functionality. To date, several clinical trials of tracheal replacement with tissue-engineered trachea have been performed. This article reviews the formation of cartilage tissue, epithelium and neovascularization of tissue-engineered trachea, together with the obstacles, possible solutions and future. Furthermore, the role of the bioreactor for in vitro tracheal graft formation and recently reported clinical applications of tracheal graft were also discussed. Generally, although encouraging results have been achieved, however, some obstacles remain to be resolved before the tissue-engineered trachea can be widely used in clinical settings.
    Matched MeSH terms: Tissue Culture Techniques
  20. 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
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