• Asian-Australasian Journal of Animal Sciences
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• v.14 no.1
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• pp.17-20
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• 2001

The effect of chicken IGF-I on protein synthesis of chicken embryo myoblasts cultured in serum-free medium was examined. When myoblasts were expanded to approximate 20-30% of well, the medium was changed to the serum-free medium including 0, 2, 20, 200 or 2000 ng/ml of recombinant chicken IGF-I. The culture medium including 10% fetal calf serum (FCS) was used as positive control. After 1 day of incubation, protein synthesis was measured by the incorporation of [$^3H$]-L-leucine. Thereafter cells were continued to incubate for further 18 hours, and the radioactivity in the protein was measured as an index of protein synthesis. The values for protein synthesis cultured in the serum-free medium without chicken IGF-I or with 2000 ng/ml of chicken IGF-I were the lowest. Protein synthesis was elevated with increasing chicken IGF-I concentration from 0 to 20 ng/ml. The values for protein synthesis in the 20 ng/ml and 200 ng/ml IGF-I groups were about half of that of the FCS group. The present study revealed that the potency of chicken IGF-I at the levels of 20 to 200 ng/ml to stimulate myoblast protein synthesis was about half of that of 10% FCS.

• Microbiology and Biotechnology Letters
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• v.17 no.3
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• pp.231-235
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• 1989

Possibility of using microcarriers for the growth of a transformed human embryonic kidney cell line 293 was investigated. The cell grew well in a static culture such as T-flasks with medium of DME/F12 (3:1) mixture supplemented with 5% FBS, but it was most difficult to make the cells grow on microcarriers mainly due to the low attachment efficiency and poor spreading at initial stage of the culture. Consequently, 30-50% of the cells were lost upon inoculation into microcarrier suspension and significant fraction of the mirrocarrier became bald. The medium supplemented with the concentrated conditioned medium by hepatoma cell line HpG2 supported the active growth of the cells on microcarrier and the cells showed a very healthy and well spreading morphology. It was probable that some spreading and attachment factors of HpG2 conditioned medium were effective for 293 cells.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.4
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• pp.319-324
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• 2006

Although the sera used in animal cell culture media provide the macromolecules, nutrients, hormones, and growth factors necessary to support cell growth, it could also be an obstacle to the production of recombinant proteins in animal cell culture systems used in many sectors of the biotechnology industry. For this reason, many research groups, including our laboratory, have been trying to develop serum-free media (SFM) or serum-supplemented media (SSM) for special or multi-purpose cell lines. The Chinese hamster ovary (CHO) cell, for example, is frequently used to produce proteins and is especially valuable in the large-scale production of pharmaceutically important proteins, yet information about its genome is lacking. Also, SFMs have only been evaluated by comparing growth patterns for cells grown in SFMs with those grown in SSM or by measuring the titer of the target protein obtained from cells grown in each type of medium. These are not reliable methods of obtaining the type of information needed to determine whether an SFM should be replaced with an SSM. We carried out a cDNA microarray analysis to evaluate MED-3, an SFM developed in our laboratory, as a CHO culture medium When CHO cells were cultured in MED-3 instead of an SSM, several genes associated with cell growth were down-regulated, although this change diminished over time. We found that the insulin-like growth factor (IGF) gene was representative of the proteins that were down-regulated in cells cultured in MED-3. When several key supplements - including insulin, transferrin, ethanolamine, and selenium - were removed from MED-3, the IGF expression was consistently down- regulated and cell growth decreased proportionately. Based on these results, we concluded that when an SFM is used as a culture medium, it is important to supplement it with substances that can help the cells maintain a high level of IGF expression. The data presented in this study, therefore, might provide useful information for the design and development of SFM or SSM, as well as for the design of genome-based studies of CHO cells to determine how they can be used optimally for protein production in pharmaceutical and biomedical research.

• KSBB Journal
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• v.13 no.2
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• pp.181-186
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• 1998

In order to obtain high-level production of recombinant human thrombopoietin (rhTPO) in insect cell line, HTI-TN-5B1-4 (TN5), conditions for optimal rhTPO expression such as multiplicity of infection (MOI), the cell density at infection, harvesting time and type of culture method as well as growth media were determined. When TN5 cells were cultured as anchorage-dependent state in 60-mm dish, cell density $2\times^6$ cells,MOI of 10 and Garvesting the culture media at 72 hr post-infection wrere the cinditions for highest rh TPO production. High production of rhTPO was also achieved by using EXPRESS FIVE serum free media rather than SF900II serum free media-1. Anchorage-dependent TN5 cells were adapted as a suspension culture when they were grown in the presence of heparin. TN5 cells were successfully cultured at 0.2 L scale in suspension culture without having aggregation. When TN5 cells were cultured as suspension state, cell density of $0.6\times10^6$ cells/mL, MOI of 1 and harvesting the culture media at 72 hr post-infection, gave the highest yield of rhTPO.

• Korean Journal of Animal Reproduction
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• v.22 no.1
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• pp.51-59
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• 1998

This study was undertaken to investigate the effects of ITS and EGF on embryonic development of in vitro matured and fertilized bovine oocytes in culture medium su, pp.ementing with or without calf serum. When fertilized oocytes were cultured in TCM-199 containing 0, 0.1, 0.5 and 1.0\ulcorner/ml ITS with 5% calf serum, the rates of development to blastocyst stage and the cell number of blastocysts were not significantly different among all treatments. And also monolayer of cumulus/granulosa cells prepared in containing calf serum and ITS were no beneficial effects of embryonic development. On the other hand, when EGF was su, pp.imented to TCM-199 containing calf serum or calf serum free, embryonic development rates(24.0 2.8% to 29.2 1.7% or 8% to 9%) and cell number of blastocysts(p<0.05) were significantly increased compared with EGF-free(22.1 2.1 or 1.0%, p<0.05). But when fertilized oocytes were cultured with cumulus/granurosa cells in TCM-199 containing EGF and calf serum, the rate of embryos development to the blastocyst stage and cell number of blastocysts were not significantly different compared with EGF-free and any concentrations. These results showed that ITS and EGF was not improved the development of bovine embryo in vitro matured and in vitro fertilized with calf serum and/or monolayer of cumulus/granulosa cells.

• 한국생물공학회:학술대회논문집
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• 2005.04a
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• pp.203-206
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• 2005

TGE (Transmissible Gastroenteritis) caused by a virus belonging to family coronavirus, results in an acute infection of the small intestine of the pig. The optimum operation variables such as multiplicity of infection (MOI), infection time and harvest time were investigated for TGE vaccine production by immobilized ST(swine testicle) cells. In the culture supplemented with 5% serum, maximum virus titer of $1.2{\times}10^6pfu/ml$ was obtained at the conditions of 0.01 MOI, 2day infection time, and 1 day harvest time. Serum is a potential source of bacterial, mycoplasmal and viral contamination, and it has a possibility of the introduction of serum proteins, prion and pyrogens into the final product. For these reasons, much attention has been focused on the development of serum-free media. A new serum-free media (SFM) has been developed in order to produce TGE vaccine of high quality with low cost. The performance of SFM developed was compared with other commercially available serum-free media and serum supplemented media in terms of virus productivity. The cultures with serum-free media showed higher titer than that with serum supplemented media. Among various serum-free media tested, CHO-S-SFMII showed highest virus titer.

• KSBB Journal
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• v.9 no.3
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• pp.272-278
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• 1994

Attenuated varicella-zoster virus(VZV) was cultured in human embryonic lung cells. The effects of serum type and its concentration on the production of VZV were studied. Regardless of cell culture conditions, VZV yield was increased with multiplicity of infection, and the total cell concentration was decreased after virus infection. The newborn calf serum, calf serum, or horse serum was not as good as the fetal bovine serum or calf serum supplemented with iron for the propagation of VZV in the human embryonic lung cells. The yields of total VZV(cell-associated virus and cell-free virus) in the medium with calf serum containing iron were comparable to those in the medium supplemented with fetal bovine serum. Furthermore, some components of serum appear to be important for the maintenance of VZV infectivity.

• The Korean Journal of Pharmacology
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• v.31 no.1 s.57
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• pp.85-93
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• 1995

In order to examine the effect of testosterone of the cell growth, using a primary rabbit kidney proximal tubule cell culture system, we observed the effect of 3 growth factors and testosterone supplementation on the growth of primary rabbit kidney proximal tubule cells in the serum-free medium. 1 nM of testosterone showed a potentiation of the effect on the growth of the proximal tubule cell in serum-free medium, but higher concentration (>10 nM) of testosterone indeed inhibited the growth. In the absence of hydrocortisone as a growth supplement in serum-free medium, testosterone caused to potentiate the growth of the cell. In the presence of hydrocortisone, testosterone also potentiated the grwoth of the proximal tubule cells. According to the Northern analysis, testosterone increased significantly the level of ${\beta}-actin$ mRNA in proximal tubular cells of rabbit kidney. Consequently we may suggest that growth stimulatory effect of testosterone on the primary rabbit kidney proximal tubule cell in serum-free and hormonally defined media ascribed to increase the synthesis of ${\beta}-actin$, which is an important protein consisting of cellular microfilament.

• 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 the Korean Society of Food Culture
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• v.23 no.4
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• pp.521-528
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• 2008

This study examined the effects of 2% Lycii fructus powder (LFP) supplementation on lipid metabolism in rats. Male Sprague-Dawley rats were administered 1% cholesterol and 0.25% sodium cholate to induce hypercholesterolemia. Then, 40 rats were divided into four diet groups: a normal diet group (NC), high cholesterol diet group (HC), normal diet plus 2% Lycii fructus powder (NC-LFP) group, and high cholesterol diet plus 2% Lycii fructus powder (HC-LFP) group. The HC group presented higher growth rates and liver weights than NC and NC-LFP however, growth rates and liver weights in the 2% LFP administered groups gradually decreased. HC also showed increased serum triglyceride, total cholesterol, free cholesterol, and LDL-cholesterol levels and decreased atherogenic index values, HDL-cholesterol, and phospholipid levels, whereas LFP group showed decreased serum triglyceride, total cholesterol, free cholesterol, and LDL-cholesterol levels as compared to HC. There were no differences in serum triglyceride, phospholipid, LDL-cholesterol, HDL-cholesterol, and free cholesterol concentrations between the normal diet groups (NC and NC-LFP). The high cholesterol diet groups (HC and HC-LFP) had significant increases in serum alanine aminotransferase (ALT), asparate aminotransferase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDHase) activities. And the 2% LFP administered groups had lower hepatic concentrations of total cholesterol and triglycerides than the HC groups. Overall, the results suggest that Lycii fructus powder has hypochloesterolemic effects by reducing serum and liver cholesterol contents.