• Title/Summary/Keyword: Cell culturing

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Comparison of Culture Media for In Vitro Maturation of Oocytes of Indigenous Zebu Cows in Bangladesh

  • Singha, Joydev Kumar;Bhuiyan, Mohammad Musharraf Uddin;Rahman, Mohammad Moshiur;Bari, Farida Yeasmin
    • Journal of Embryo Transfer
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    • v.30 no.4
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    • pp.327-333
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    • 2015
  • The objectives of the present study were to select an effective basic medium including its hormone and protein supplementation for IVM of oocytes of indigenous zebu cows. The ovaries of cows were collected from slaughter house and the follicular fluid was aspirated from 2 to 8 mm diameter follicles. The COCs with more than 3 cumulus cell layers and homogenous cytoplasm were selected for maturation. The oocytes were matured in media for 24 hrs at $39^{\circ}C$ with 5% $CO_2$ in humidified air. The maturation of oocytes was evaluated by examining the presence of first polar body under microscope. An efficient basic medium was determined after culturing COCs in either TCM 199 or SOF medium in Experiment 1. An efficient hormone supplementation was determined after culturing COCs in either FSH or gonadotrophin supplemented TCM 199 in Experiment 2. An efficient protein supplementation was determined after culturing COCs in either FBS or Oestrous cow serum (OCS) supplemented TCM 199 in Experiment 3. The oocyte recovery rate per ovary was 3.35. The overall rate of IVM was 74.6%. The maturation rate was $75.5{\pm}3.9$ and $62.2{\pm}20.2%$ in TCM and SOF medium, respectively (P>0.05). The maturation rate of oocytes was significantly higher ($76.6{\pm}13.2%$) in FSH supplemented medium than gonadotrphin supplemented counterpart ($69.7{\pm}10.8%$) (P<0.05). The maturation rates of oocytes were $81.7{\pm}12.9$ and $85.7{\pm}12.7%$ in medium supplemented with FBS and OCS, respectively (P>0.05). In conclusions, both TCM 199 and SOF supplemented with either FBS or OCS, and FSH may be used as medium for IVM of indigenous zebu oocytes in Bangladesh.

Studies on Sexing of Bisected Mouse Embryos by Rat H-Y Antibody (Rat H-Y 항체에 의한 생쥐 분할란의 성 조절에 관한 연구)

  • 정장용;박희성;박충생
    • Korean Journal of Animal Reproduction
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    • v.15 no.3
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    • pp.179-187
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    • 1991
  • This experiment was carried out to develop a new technique of identifying XX of XY-bearing bisected embryos prior to implantation by immunological method. H-Y antiserum prepared in inbred Wastar female rats by repeated immunization with spleen cells from males of the same strain. The reactivity of H-Y antibody was confirmed by culturing mouse embryos in the medium containing H-Y antiserum and complement obtained from the guinea pig. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos under the concentraton or affected H-Y antibody was also investigated by culturing embryos under the concentration or affected H-Y antibody and culture rate. However, production of live young or sex rates of male and female from embryos transferred with psudopregnant. The biological test with the morula stage embryos showed that H-Y antibody was formed in all female rats immunized with spleen cell, but it was formed only in 80% female rats immunized with the antigen. When the bisected mouse embryos were cultured in vitro for 5~6 hours in morula stage, of 457 bisected embryos 81.4% of then were developed to the blastocyst stage. When the concentration rate of complement to H-Y antiserum varied from 1.0~5.0${mu}ell$, the lysis-rate of embryo was 19.5 to 67.3%. The concentration rate of complement did not influence the lysis-rate of embryos(P<0.05). The morphology embryos of bisected, zona-free and intact embryos showed the embryos lysis rate of 58.6, 42.7 and 48.5% respectively(P<0.05). Pregnancy rate were 50.0, 45.5 and 57.1% in psudopregnant recipient transferred with bisected, zona-free and intact blastocyst embryos. However, production of live youngs, sexual rate of male or female was 24(50.0:50.0), 22(45.5:55.5) and 36(58.3:41.7)mice, but affected and non affected half embryos with H-Y antiserum treatment was 23.1 and 26.7%. Also production of live youngs and sexual rate was 14(92.9:7.1) and 17(17.6:82.4)mice in affected and non affected half embryos in H-Y antiserum treatment(P<0.05).

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Preparation and Characterization of Genetically Engineered Mesenchymal Stem Cell Aggregates for Regenerative Medicine

  • Kim, Sun-Hwa;Moon, Hyung-Ho;Chung, Bong-Genn;Choi, Dong-Hoon
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.333-337
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    • 2010
  • Combining cell- and gene-based therapy is a promising therapeutic strategy in regenerative medicine. The aim of this study was to develop genetically modified mesenchymal stem cell (MSC) aggregates using a poly(ethylene glycol) (PEG) hydrogel micro-well array technique. Stable PEG hydrogel micro-well arrays with diameters of 200 to $500\;{\mu}m$ were fabricated and used to generate genetically engineered MSC aggregates. Rat bone marrow-derived MSCs were transfected with a green fluorescent protein (GFP) plasmid as a reporter gene, and aggregated by culturing in the PEG hydrogel micro-well arrays. The resultant cell aggregates had a mean diameter of less than $200\;{\mu}m$, and maintained the mesenchymal phenotype even after genetic modification and cell aggregation. Transplantation of MSC aggregates that are genetically modified to express therapeutic or cell-survival genes may be a potential therapeutic approach for regenerative medicine.

Production of Volatile Oil Components by Cell Culture of Agastache rugosa O. Kuntze

  • Shin, Seung-Won;Kim, You-Sun;Kang, Chan-Ah
    • Natural Product Sciences
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    • v.7 no.4
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    • pp.120-123
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    • 2001
  • To develop systems for economic production of useful essential oil compounds, callus was induced from the seedlings of Agastache rugosa and cultured on MS medium. The volatile oil fraction was extracted from the callus and investigated by mean of GC-MS. The composition of the oil was compared with that of the mother plant. As a result, sixty five compounds including ferruginol were identified in the essential oil fraction. The main component of the oil from the leaves of Agastache rugosa was methyl chavichol (53.6%). Methyl jasmonate and jasmonic acid were added to the culturing cell suspension, separately and the composition of induced oil were compared. The oils from cultured cells treated with jasmonates showed considerably different patterns. Especially, the peak of estragole was found in callus oil after treatment with methyl jasmonate as though the amount was limited to 0.58%. In general, the TIC pattern of GC-MS of the callus oil became more similar to the oil from the leaves after elicitation.

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Influence of Organic Carbon Sources on Growth and Lipid Content of Marine Green Alga Dunaliella tertiolecta

  • Rizwan, Muhammad;Mujtaba, Ghulam;Lee, Kisay
    • Journal of Marine Bioscience and Biotechnology
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    • v.6 no.2
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    • pp.68-75
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    • 2014
  • This study investigated the potential use of various organic carbon sources (glucose, glycerol and acetate) and different concentrations of $CO_2$ for culturing marine microalga Dunaliella tertiolecta. Cell growth and lipid production were monitored under heterotrophic, mixotrophic and photoautotrophic modes of cultivation. D. tertiolecta showed the ability to grow under mixotrophic (acetate and glucose), heterotrophic (glucose) and photoautotrophic condition under high $CO_2$ concentration (15%). With all the organic carbon sources (glucose, glycerol and acetate) tested in this study, 1~5% acetate enhanced cell growth rate and lipid content, while higher concentrations of acetate (10% and 15%) were inhibitory and resulted in cell death.

Dental Pulp Stem Cell: A review of factors that influence the therapeutic potential of stem cell isolates

  • Young, Aubrey;Kingsley, Karl
    • Biomaterials and Biomechanics in Bioengineering
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    • v.2 no.2
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    • pp.61-69
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    • 2015
  • Undifferentiated stem cells are being studied to obtain information on the therapeutic potential of isolates that are produced. Dental Pulp Stem Ccell (DPSC) may provide an abundant supply of highly proliferative, multipotent Mesenchymal Stem Cells (MSC), which are now known to be capable of regenerating a variety of human tissues including bone and other dental structures. Many factors influence DPSC quality and quantity, including the specific methods used to isolate, collect, concentrate, and store these isolates once they are removed. Ancillary factors, such as the choice of media, the selection of early versus late passage cells, and cryopreservation techniques may also influence the differentiation potential and proliferative capacity of DPSC isolates. This literature review concludes that due to the delicate nature of DPSC, more research is needed for dental researchers and clinicians to more fully explore the feasibility and potential for isolating and culturing DPSCs extracted from adult human teeth in order to provide more accurate and informed advice for this newly developing field of regenerative medicine.

Optimal Milieu for Culturing Porcine Sertoli Cell

  • Jabed Md. Anower;Kamal Tania;Kim, Byung-Ki
    • Reproductive and Developmental Biology
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    • v.30 no.3
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    • pp.163-167
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    • 2006
  • The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

Recent Trends of the Development of Photobioreactors to Cultivate Microalgae (미세조류 배양을 위한 광생물반응기 개발의 기술동향)

  • Kim, Jong-Tye;Ahn, Dong-Gyu;Park, Jong-Rak;Park, Jeong-Woo;Jeong, Sang-Hwa
    • Journal of the Korean Society for Precision Engineering
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    • v.28 no.2
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    • pp.125-132
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    • 2011
  • This paper describes current status and future prospects of the mass production of microalgae biomass. Microalgae have attracted considerable attention since they not only effectively fix $CO_2$ gas during their metabolic process but also have the great potential to be utilized for producing valuable substances as a kind of efficient light-harvesting cell factories. In this review, we outline various types of photobioreactors employed for mass production of biomass by culturing microalgae in a well controlled way and give an overview about the present state of affairs, both domestic and international, in the field of the microalgal culturing technologies.

Properties of BE0623 to serve as a growth factor of Bifidobacterium

  • Cho, Young Hoon;Sim, Jae Young;Nam, Myoung Soo
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.445-457
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    • 2020
  • Prebiotics are defined as "Nondigestible food ingredients that beneficially affect the host by selectively stimulating the growth and activity of bacteria in the intestine" and as defined improve host health. This study was carried out to investigate the effects of bifidobacteria (Bifidobacterium lactis BB12 and Bifidobacterium longum BB536) growth enhancer (BE0623) supplement as a prebiotic. The addition of BE0623, a growth promoting material for bifidobacteria, significantly increased bifidobacteria viable cells counts in fermented milk by about 45 to 75 times compared to the non-added control group. In addition, microscopic observation showed a significant effect on proliferation of bifidobacteria in fermented milk with added BE0623. The viable cell counts in bifidobacteria also increased roughly 102-fold compared to the control group (non-added BE0623) and was higher than that of commercial growth promoters. Each fraction obtained though the purification of BE0623 influenced the increase of bifidobacteria growth. Culturing bifidobacteria with a combination of fractions of BE0623 had a synergistic effect compared to culturing bifidobacteria with each fraction individually. When any of the fractions were not added, the effect of the growth enhancer on bifidobacteria was reduced. These results indicate that all fractions contain substances that promote the growth of bifidobacteria. Therefore, BE0623 is considered to be available as a growth promoting material for bifidobacterium.

Optimal Condition and Interspecific Cross-Reaction of H-Y Antibody Activity (H-Y항체활성의 최적조건과 종간교차반응)

  • ;H.S.Shim;J.B.Kim;H.Y.Park;K.S.Chung
    • Korean Journal of Animal Reproduction
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    • v.10 no.2
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    • pp.168-174
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    • 1986
  • These experiments were carried out to clarify the optimalconditions and interspecific cross reaction of H-Y antibody activity. H-Y antiserum was prepared in inbred SD female rats and Balb/c female mice by repeated immunization of rat newborn testis homogemate, rat and mouse spleen cells obtained from males of same strain. The activity of H-Y antibody in antiserum was tested by ELISA and biological tests. The cross reactivity of H-Y antibody was confirmed by culturing mouse and rabbit embryos in medium containing H-Y antibody and complement obtained from rat and guinea pig, respectively. The optimal condition for the activity of H-Y antibody was also investigated by culturing embryos in medium with different pH and complement concentration. The results obtained in these experiments were summarized as follows: 1. The formation rates of H-Y antibody in rats immunized with newborn testis and spleen cell were 40.0 and 50.0% respectively, and that in mouse immunized with spleen cell was 48.4%. 2. The activity of H-Y antibody was not affected by pH in range of 6.5 to 8.0, and the same was true for the relative concentration of complement to the H-Y antibody. 3. Minimum time needed for the activity of H-Y antibody was confirmed to be 0.5 to 1 hour and 24 to 48 hours respectively for the zona free embryos and intact embryos. 4. When mouse and rabbit embryos were treated with H-Y antibody obtained from rat, 46.4 and 54.8% of embryos were retarded or destroyed. From these results it could be said that H-Y antibody had strong interspecific cross reactivity.

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