• Title/Summary/Keyword: Fetal bovine serum

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Enhanced Expression of Foreign Gene in Baculovirus-Infected Insect Cells Using a Silkworm Hemolymph (누에 체액을 이용한 외래 유전자의 발현효율 증대)

  • 우수동;김혜성
    • Journal of Sericultural and Entomological Science
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    • v.37 no.2
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    • pp.181-185
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    • 1995
  • To enhance expression of foreign gene by the novel expression vector, pBmKSK1, of Bombyx mori nuclear polyhedrosis virus, E. coli $\beta$-galactosidase gene expressing recombinant virus was infected in BmN-4 cells and various concentrations of silkworm hemolymph were added to the recombinant virus-infected BmN-4 cells containing fetal bovine serum. The expression efficiency of foreign gene was determined by $\beta$-galactosidase activity in the culture media. The results showed that the silkworm hemolymph was effective to expression of foreign gene in the BmN-4 cells, suggesting that the silkworm hemolymph could be substituted for fetal bovine serum in the BmN-4 cells to enhance expression of foreign gene.

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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.

Effect of Blood Contamination on Vickers Microhardness and Surface Morphology of Mineral Trioxide Aggregate

  • Jaehyun Seung;Seong-Jin Shin;Byounghwa Kim;Ji-Myung Bae;Jiyoung Ra
    • Journal of the korean academy of Pediatric Dentistry
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    • v.51 no.2
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    • pp.165-175
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    • 2024
  • This study aimed to investigate the effects of blood contamination on the Vickers hardness and the surface morphology of premixed MTA and compare them with the effects on conventional MTA. The Vickers microhardness of Endocem MTA Premixed Regular (EP) and ProRoot MTA (PM) was assessed after immersion in fetal bovine serum (FBS) and saline. Stem cells from human exfoliated deciduous teeth (SHED) were seeded on MTA after immersion in FBS, saline, and deionized water (DW). Cell adhesion patterns and surface morphology were visualized via scanning electron microscopy (SEM). The surface microhardness of EP and PM in FBS was lower than in saline. However, short-term exposure of PM to FBS did not reduce the microhardness compared to saline. Angular crystals formed in water, while rounded crystals with more air voids appeared in FBS. Favorable SHED attachment occurred in all groups. Overall, the surface hardness of EP and PM decreased after FBS exposure, although PM was less influenced. We suggest minimizing the amount of bleeding when using MTA clinically; nevertheless, PM remains an option with more expected blood contamination than EP. In summary, exposure to FBS decreased mechanical performance but allowed cell adhesion for both MTAs, with PM being more resistant to these changes.

Bovine Nuclear Transfer using Ear Skin Fibroblast Cells Derived from Serum Starvation and Passage Numbers

  • Yang, Byoung-Chul;Im, Gi-Sun;Park, Jin-Ki;Kim, Hyun-Ju;Chang, Won-Kyung
    • Proceedings of the KSAR Conference
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    • 2001.03a
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    • pp.64-64
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    • 2001
  • To facilitate the widespread application of somatic cell cloning, improvements in blastocyst production efficiency and subsequent fetal viability are required. Area where technical improvements are needed include donor cell treatments, starvation and passage numbers. This study was carried out to investigate the effect of serum-starvation and passage on the development of ear skin fibroblast cells cloned embryos. A skin biopsy was obtained from the ear of a 2-year-old Korean Hanwoo female. The cells were cultured in 10% FBS+DMEM up to 2-3 months(up to 10 passages) and then used. In Experiment 1, the Korean bovine Ear Skin Fibroblast cells (KbESF) were either serum starved (culture in 0.05% FBS+DMEM) or serum fed (10% FBS+DMEM) for 4-7 days Prior to NT In Experiment 2, the KbESF cells used for nuclear transfer in these experiments were from passages 2 to 10. The development of 208 nuclear transfer (NT) embryos reconstructed from either serum starved or serum fed ear skin fibroblast was assessed. NT embryos reconstructed from serum starved and serum fed cells showed the same developmental rate (cleavage 80.16 vs. 85.37%; blastocyst 20.63 vs. 19,51%). The development of 590 nuclear transfer (NT) embryos reconstructed from passage 2 to 10 was assessed. We observed the same developmental rates for embryos derived from later Passages as compared with those embryos from early passages(blastocyst from 16.69 to 27.91%, average 20.17%). There was no significant difference between serum-fed and serum-starved donor cells. We observed no difference in developmental rates for embryos derived from 2 to 10 passages. These data show that prolonged culture and serum starvation does not affects the cloning competence of adult somatic cells.

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Phytomitogen Induced Blast Transformation of Guinea Pig: I. Effect of Concentration of Serum and Time of Incubation on the Uptake of 3H -Thymidine (Phytomitogen에 의한 기니픽 임파구의 Blast Transformation: II. 유사분열에 미치는 혈청농도와 배양시간의 효과)

  • Kim, Jong-shu
    • Korean Journal of Veterinary Research
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    • v.27 no.1
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    • pp.41-45
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    • 1987
  • The present study was carried out to investigate the optimal conditions on blastogenesis of guinea pig lymphocytes. A microculture system in conjuction with a semiautomatic multiple sample harvester was used to study the in vitro optimal condition of guinea pig lymphocytes. Careful analysis of lymphocyte transformation to concanavalin A (Con A) and lipopolysaccharide (LPS) mitogen determined optimal conditions as: (a) 10% fetal bovine serum in RPMI-1640 medium (b) 48-hour culture period.

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Inhibition of Oral Epithelial Cell Growth in vitro by Epigallocatechin-3-gallate; Its Modulation by Serum and Antioxidant Enzymes

  • Hong, Jung-Il;Kim, Mi-Ri;Lee, Na-Hyun;Lee, Bo-Hyun
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.971-977
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    • 2009
  • The most abundant tea catechin, epigallocatechin-3-gallate (EGCG), has been reported to inhibit cell proliferation and induce apoptosis in many types of cancer cells. In the present study, effects of EGCG on the growth of oral epithelial cells including CAL-27 oral squamous carcinoma cells and dysplastic oral keratinocytes (DOK) were investigated. EGCG inhibited growth of CAL-27 cells and DOK with $IC_{50}$ of 14.4-21.0 and 5.8-14.2 ${\mu}M$ after 24 and 48 hr incubation, respectively. EGCG was significantly less effective in inhibiting DOK growth. The effects of EGCG, however, were dramatically less pronounced in the presence of superoxide dismutase (SOD) and catalase. Inhibitory effects of EGCG on CAL-27 cell growth were also much less pronounced in the presence of fetal bovine serum (FBS). EGCG induced caspase-3 activation in both CAL-27 and DOK cells in a serum free condition without SOD/catalase; in the presence of 10% FBS and SOD/catalase, EGCG, even at 100 ${\mu}M$, did not affect cell growth. The present results indicate that EGCG inhibited oral cell growth with higher potency to more malignant CAL-27 cells than DOK, and the effects were markedly altered by SOD/catalase and serum content in media.

Effects of Progesterone and 17β-Estradiol under Presence or Absence of FBS on Plasminogen Activators Activity in Porcine Uterine Epithelial Cells

  • Hwangbo, Yong;Lee, Mi-Rim;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.22 no.4
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    • pp.309-318
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    • 2018
  • The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) activation by $17{\beta}$-estradiol ($E_2$) and progesterone ($P_4$) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL $E_2$, and $P_4$ with or without fetal bovine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and expression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of $E_2$ compared to 0.1% DMSO treatment in serum-free group (p<0.05). Then, $E_2$ and $P_4$ were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cultured cells without FBS. Only tPA mRNA was significantly increased by $E_2$ treatment (p<0.05). PAs activity was enhanced in $E_2$ treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to evaluate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.

Effect of Gender-specific Bovine Serum Supplemented Medium on Cell Culture (성별 특이 소 혈청이 세포 배양에 미치는 영향)

  • Lee, Dong-Mok;Choi, Moon-Seok;Woo, Gyung-Il;Shin, Yu-Mi;Lee, Ki-Ho;Cheon, Yong-Pil;Chun, Tae-Hoon;Choi, In-Ho
    • Journal of Animal Science and Technology
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    • v.51 no.5
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    • pp.413-420
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    • 2009
  • The research has been aimed to investigate the effect of different sera including fetal bovine serum (FBS), male bovine serum (MS), female bovine serum (FS), and castrated-male bovine serum (C-MS) on cell proliferation, follicular maturation and ovulation in vitro. Established cell lines and primary cells were cultured in the culture media supplemented with different sera and cells proliferation was observed by cell counting and MTT assay. The results indicated that cell proliferation was significantly different for different serum source. Proliferation of bovine and human myogenic satellite cells was highest in MS. In contrast, proliferation of breast cancer cells and immune cells were the highest in FS and FBS, respectively. There was no difference in the rate of follicular growth, whereas the rate of ovulation was higher in FBS and C-MS. Finally, the wound healing effect and cell proliferation of 3T3-L1 cells showed that wound healing was fastest in FS and cell proliferation was higher in MS. These results suggest the importance of an optimal serum selection in the experiments involving cell culture system, and gender-specific Hanwoo sera may be used as a substitute to FBS.

Effect of Quiescent Treatment on Nuclear Remodeling and In Vitro Development of Nuclear Transfer Embryos Derived from Bovine Fetal Fibroblast Cells (세포의 휴면처리가 소 태아섬유아세포 유래 핵이식란의 핵상변화와 체외발육에 미치는 영향)

  • 최종엽;권대진;김정익;박춘근;양부근;정희태
    • Korean Journal of Animal Reproduction
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    • v.24 no.2
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    • pp.217-222
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    • 2000
  • This study was conducted to investigate the effect of quiescent treatment of the donor cells on the nuclear remodeling and in vitro development of fetal fibroblast cell-cloned bovine embryos. Serum starved, confluent and nonquiescent cycling fetal fibroblast cells were transferred into the enucleated oocytes. About 20∼25% of nuclear transfer embryos fused with a serum starved or confluent cell extruded a polar body, which was slightly lower than that of nontreated control (36%). About 49∼51% of nuclear transfer embryos fused with a serum starved or confluent cell had a single chromatin clump, which was slightly higher than that of nontreated control (40%). The proportion of embryos with a single chromatin clump was significantly higher (P<0.01) in nuclear transfer embryos without showing a polar body (60.5%) than with a polar body (4.7%). Development rates to the blastocyst stage were 21.7% and 20.9% when serum starved and confluent cells were transferred, which were slightly higher than that of control (14.1 %). The result of this study suggests that quiescent treatment by serum starvation or growth to confluency of donor cells could increase the number of embryos with a normal chromatin structure, which results in increased in vitro development.

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