• Journal of Life Science
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• v.24 no.12
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• pp.1301-1307
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• 2014

Adipose-derived stem cells (ADSCs) are considered promising tools for tissue regeneration. However, ADSCs have very poor proliferation capacity. Therefore, fetal bovine serum (FBS) is generally added to the culture media of ADSCs. As FBS contains many uncharacterized components that may affect cellular functions, methods for serum-free cultures of ADSCs have been widely investigated. In this study, to develop an efficient method for a serum-free culture of ADSC-T, we used an ADSC line established by introducing the simian virus 40 (SV40) T gene into primary ADSCs. We then investigated the effect of amino acids, vitamins, and other components on the growth of ADSC-T. When the ADSC-T cells were plated with DMEM/F12 serum-free medium, the cells did not proliferate, and the mixture of amino acids, vitamins, and B27 supplement did not increase the growth of the cells. However, when the ADSC-T cells were provided with serum-free DMEM/F12 after they had been cultured with serum-supplemented DMEM for 24 h, the cells proliferated, and the vitamins and B27 supplement increased the cell growth. Stem-Pro serum-free medium also appeared to be useful as a suspension culture for the ADSC-T cells. The ADSC-T cells secreted large amounts of proteins of around 70 kDa. Insulin-like growth factor (IGF) and fibroblast growth factor basic (FGF basic) were secreted by ADSC-T in larger amounts in the serum-free culture than in the serum-supplemented culture.

• KSBB Journal
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• v.5 no.1
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• pp.87-94
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• 1990

Enhancement of hybridoma cell growth and monoclonal antibody(MAb) production by the addition of a small amount of serum into both serum-free medium and enriched medium was studied. The enriched medium was constructed by mixing a basal serum-free medium and a nutrient-fortified RPMI 1640 medium. It was supplemented with human serum albumin, insulin, transferrin, and monoethanolamine. It was found that addition of low concentration of serum with other serum-free supplements was favorable for growth of a mouse hybridoma 2c3.1 cells. The concentration of serum was determined to 0.5%. The maximum cell concentration obtained in this enriched medium supplemented with 0.5% fetal bovine serum (FBS) was $3.06{\times}10^6$ cells/ml and the concentration of secreted anti-Hepatitis surface antigen (antiHBsAg) MAb was $159.7{\mu\textrm{g}}\;/\;ml$ compared to $43{\mu\textrm{g}}\;/\;ml$ in RPMI 1640 medium with 10% FBS and $50{\mu\textrm{g}}\;/\;ml$ in previously-developed serum-free medium. The 2c3.1 cell growth and MAb production could be enhanced considerably by using the enriched medium supplemented with 0.5% FBS and serum-free supplements instead of RPMI 1640 medium or serum-free medium. The enhancement in MAb production in the enriched medium was more noticeable.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.108-108
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• 2005

• Journal of Embryo Transfer
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• v.24 no.4
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• pp.281-287
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• 2009

As a simple and economical method for in vitro produced embryos, we have used BSA instead of serum for the production and embryo transfer of Hanwoo in vitro fertilized (IVF) embryos and obtained the following results: 1) When using serum (FBS; fetal bovine serum) or BSA-containing culture media as the initial culture media for immature oocytes, it is regarded as inappropriate to add only BSA to the culture solutions from maturation of the immature oocytes to development stage culture, but serum still needs be added though there is no significant difference in the concentration, with a change from 5% to 10%. 2) The results of culturing IVF embryos after development (4 cell stage) in the Medium199 solutions containing BSA instead of serum (FBS) showed that 0.3% BSA concentration is not optimal and 0.5% or higher BSA concentration has no significant difference among 0.5%, 0.7%, 1% and 2% (p > 0.05). 3) The post-freezing survival ratio after development in 5% FBS-Medium199 showed that 1% BSA concentration of the culture solution is the most suitable in the BSA concentrations of 0.3% (51%), 0.5% (67%), 0.7% (69%), 1% (77%) and 2% (75%). 4) The pregnancy rates of the transplanted fresh(not frozen) blastocyst had no significant concentration dependency (p > 0.5), and the average pregnancy rate was 63.8%. 14% of overweight calves were found among the calves given birth to by the transfer of IVF blastocysts cultured in the serum-added culture solution, but none was found in the experimental groups in which BSA was added instead of serum.

• Journal of Microbiology
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• v.42 no.1
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• pp.25-31
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• 2004

Sf9 cells have obvious advantages for the conventional production technology of vaccine. They are useful tools for high concentration and large-scale cultures. Sf9 cells were grown to maximal concentration, 8${\times}$l0$\^$6/ cells/$m\ell$ in a 500$m\ell$ spinner flask, with a doubling time at the exponentially growing phase of 24.5 hours, using serum-free media. To explore the ability of Sf9 cells to be infected by the Japanese encephalitis (JE) virus Beijing-l strain, Sf9 cells were infected with the virus. By 4-5 days post-infection, 10-15 % of the Sf9 cells showed cytopathic effect (CPE), from granularity to the formation of syncytia and multinucleated giant cells continuously observed over a period of 35 days. Positive fluorescent reactions were detected in 30-40% of cells infected with the JE virus Beijing-l strain, and the uninfected Sf9 cells were completely negative. Virus particles, propagated in Sf9 and Vero cells, were concentrated by sedimentation on 40% trehalose cushions by ultracentrifugation, and showed identical patterns of viral morphogenesis. Complete virus particles, 40 to 50 nm in diameter, were observed, and JE virus envelope (E) proteins, at 53 kDa, were found in the western blot analysis to the anti-JE virus E protein monoclonal antibody and reacted as a magenta band in the same position to the glycoprotein staining. To evaluate whether the infectious virus was produced in Sf9 cells inoculated with the JE virus Beijing-l stain, Sf9 cells were inoculated with the virus, and sample harvested every 5 days. The titers of the JE virus Beijing-l strain rose from 1.0${\times}$l0$\^$5/ to 1.5${\times}$l0$\^$6/ pfu/$m\ell$. The infected Sf9 cells could be subcultured in serum-free medium, with no change in the plaque sizes formed by the JE virus Beijing-l strain in the plaque assay. It is suggested that the ability of the JE virus Beijing-l strain to infect Sf9 cells in serum-free media will provide a useful insect cell system, where the JE virus replication, cytopathogenicity and vaccine immunogen can be studied.

• KSBB Journal
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• v.21 no.2
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• pp.110-117
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• 2006

During routine maintenance, animal cell lines are commonly cryopreserved in growth medium containing serum with 10% DMSO. But, in case of bioprocess under the serum-free conditions, including cultivation of cell lines and producing of pharmaceuticals, the cryopreservation should be executed without serum to prevent a cross-contamination. This experiments were performed to investigate the effects of the serum-free cryopreservation on the CHO cells. To improve the survival rates of the cryopreserved CHO cells in serum-free condition, first, the effects of permeable and non-permeable additives for substitute serum on cell viability were investigated. The combination of 10% DMSO and 0.03 M raffinose in MEM-${\alpha}$ without serum indicated 76% of cell viability. However, it did not reach the survival rates(more than 95%) of the conventional cryopreservation. In the second, to evaluate the cryopreservative ability of the serum-free medium(SFM) we compared viability of the CHO cells cryopreserved in the SFMs(Sigma C5467, C4726, and C1707, JBI SF486 and PF486), the cryoprotectant(Genenmed CAN-1000) and the MEM-${\alpha}$ with serum. All solution contained 10% DMSO. As a result of the comparison, cryopreserved cells in the SFMs showed over 95% of viability and appeared predominant viability better than cryoprotectant CAN-1000. Finally, we assessed the stability of the CHO cells in the long-term cryopreservation(LTC) using SFM. Every three months, the cryopreserved CHO cells were thawed to estimate the cell viability and the recovery rates. Then, real-time RT-PCR analyzed the inserted CHO DHFR gene. All results for the LTC appeared the same stability as the serum containing cryopreservation. In the conclusion, it could be seen that the LTC in the SFM can substitute for serum using methods in the bioprocess proceeded by CHO cells for more than 18 months.

• Parasites, Hosts and Diseases
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• v.36 no.2
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• pp.99-108
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• 1998

The present study was performed to check the viability of eggs, filariform larvae and adults of Strongvloines venezueLensis exposed to various conditions for an in vitro maintenance. The eggs in the feces remained viable for about 25 days at $4^{\circ}C$ and 15 days at room temperature. However, the isolated eggs in sterile saline lost their viability within 24 hr at $4^{\circ}C$. The eggs in morula stage were very sensitive to air drying and rapidly lost their viability (=12 hrs. Filariform larvae survived for a maximum period of 45 days in fecal suspension and 28 days in 0.12% nutrient broth in polyvinyl culture bags maintained at $20^{\circ}C$. On the other hand, those isolated from nutrient broth cultures survived for a maximum period of 32 days in tap water and 22 days in sterile saline at $20^{\circ}C$. The mature adult worms obtained from experimentally infected rats survived maximally for 9 days in serum supplemented (10% rat-serum) 0.12% nutrient broth and 4 days in serum free nutrient broth at $37^{\circ}C$ while the culture media were changed at an alternate day. The adult female worms deposited fertile eggs in serum supplemented and serum free nutrient broth cultures, however, the hatched larvae (Ll) were not able to develop to the filariform stage in the culture media and found to die within 24 hr of maintenance. The present findings on an in vitro maintenance of different stages of 5. uenezueLetis may provide useful information for biological and biochemical studies with Strongyloines species. Key words: Strongvloides venezuelensis. viability in vitro maintenance, free-living filariform larvae (L3), embryonation of eggs

• KSBB Journal
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• v.5 no.4
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• pp.365-371
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• 1990

To improve the growth of mouse hybridoma 2c3.1 secreting anti-Hepatitis B surface antigen monoclonal antibody (anti-HBsAg MAb), we had constructed an enriched medium and observed the effects of fetal bovine serum and serum-free supplements including human serum albumin, 'insulin and transferrin', and monoethanolamine. For further enhancement of growth, the concentrations of two major energy sources, glucose and glutamine, were strengthened with various ratios in the enriched medium. Maximum cell growth and monoclonal antibody production obtained in various ratios of glucose/glutamine with an inoculation concentration of 2$\times$105 cells/ml were 0.73$\times$106-4.62$\times$106 cells/ml and 65.1-422.6 $\mu\textrm{g}$/ml, respectively. Glutamine was round to be a major energy source and a limiting nutrient in comparison to glucose for 2c3.1 cell cultivation in enriched media with low serum.

• The Journal of Internal Korean Medicine
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• v.19 no.2
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• pp.392-410
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• 1998

In order to investigate the effect of Alismatis Lhizoma on hyperlipidemia, experimental studies were performed on hyperlipidemia rats. Hyperlipidemia model (controll group) was induced by 1% cholesterol fed-diet for 8 weeks. Sample I group fed with 1% cholesterol and 4% Alismatis Lhizoma diet for 8 weeks. Sample II group fed with 1% cholesterol and 8% Alismatis Lhizoma diet for 8 weeks. The contents of serum total cholesterol, triglyceride, free fatty acid, phospholipid, HDL-cholesterol and LDL-cholesterol were measured, and fat accumulation in liver and the change of elastic and collagenous fiber in aortic wall were observed. The results were summurized as follows ; 1. The content of total cholesterol in the serum compared with control group tended to be decreased in sample group, and then sample I group showed a significant value. 2. The content of triglyceride in the serum compared with control group tended to be decreased in sample group, and then sample I group showed a significant value. 3. The content of free fat acid in the serum compared with control group tended to be decreased in sample group, but did not show a significance. 4. The content of phospholipid in the serum compared with control group tended to be decreased in sample group, and then sample I group showed a significant value. 5. The content of HDL-cholesterol in the serum compared with control group tended to be increased in sample group, and then sample II group showed a significant value. 6. The content of LDL-cholesterol in the serum compared with control group tended to be decreased in sample group, and then sample I group showed a significant value. 7. The lipophagy in liver compared with control group tended to be repressed in sample group. 8. The change of elastic and collagenous fiber lesion in tunica media of aortic wall, compared with control group tended to be repressed in sample group. According to the above results, it is assumed that Alismatis Lhizoma has a valid effect on hyperlipidemia. And yet, it needs to make further researches that sample I group showed more significant value than sample II group.

• The Journal of Internal Korean Medicine
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• v.19 no.2
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• pp.347-366
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• 1998

In order to investigate the effect of Lycii Radicis Cortex on hyperlipidemia, experimental studies were performed on hyperlipidemia rats. Hyperlipidemia model (controll group) was induced by 1% cholesterol fed-diet for 8 weeks. Sample I group fed with 1% cholesterol and 4% Lycii Radicis Cortex diet for 8 weeks. Sample II group fed with 1% cholesterol and 8% Lycii Radicis Cortex diet for 8 weeks. The contents of serum total cholesterol, triglyceride, free fatty acid, phospholipid, HDL-cholesterol and LDL-cholesterol were measured, and fat accumulation in liver and the change of elastic and collagenous fiber in aortic wall were observed. The results were summurized as follows ; 1. The content of total cholesterol in the serum compared with control group tended to be decreased in sample group, but did not show a significance. 2. The content of triglyceride in the serum compared with control group tended to be decreased in sample group, and then sample II group showed a significant value. 3. The content of free fat acid in the serum compared with control group tended to be decreased in sample group, and then sample II group showed a significant value. 4. The content of phospholipid in the serum compared with control group tended to be decreased in sample group, but did not show a significance. 5. The content of HDL-cholesterol in the serum compared with control group tended to be increased in sample group, and then sample I group showed a significant value. 6. The content of LDL-cholesterol in the serum compared with control group tended to be decreased in sample group, and then sample I group showed a significant value. 7. The lipophagy in liver compared with control group tended to be decreased in sample group. 8. The change of elastic and collagenous fiber lesion in tunica media of aortic wall, compared with control group tended to be decreased in sample group. According to the above results, it is assumed that Lycii Radicis Cortex has a valid effect on hyperlipidemia. Therefore, it seems to be applicable to the diseases related to hyperlipidemia.