• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2003년도 생물공학의 동향(XIII)
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• pp.335-339
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• 2003

본 연구에서는 연골조직으로부터 세포를 분리한 후 10%의 혈청 첨가 배지와 무혈청 배지에서의 세포 성장속도, GAGs 합성 및 Col. II의 발현을 확인하였다. EGF를 첨가한 무혈청 배지를 연골세포의 증식에 매우 효과적이었으나 GAGs 합성 및 Col. II 의 합성을 저해하였다. 또한 무혈청 배지에서 배양한 세포를 차후 신체내로 이식한다면 연골세포의 특성인 Col. II를 재합성할 수 있음을 간접적으로 확인하였다. 이는 무혈청 배지를 이용하여 평판배양 시 짧은 기간 내 연골세포 치료제로서 가능한 세포수를 얻기 위한 모델로서 유용할 것으로 생각된다. 또한 계대배양에 따른 분화 및 형태의 변화 등의 문제점들은 생분해성 지지체와 연계하여 해결할 수 있을 것으로 생각되며, 차후 위의 SFM을 이용한 3D 배양에 대한 연구를 수행할 예정이다.

• Journal of Microbiology and Biotechnology
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• 제15권6호
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• pp.1299-1303
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• 2005

Human articular chondrocytes (HAC) were cultivated as a monolayer in a serum-free medium for primary culture (SFM-P). An optimized SFM-P provides $95\%$ proliferation rate of that obtainable from primary and secondary chondrocyte cultures grown in a control medium with serum. The gradual decrease in the amounts of synthesized glycosaminoglycan and type II collagen was improved by coating the culture dishes with type IV collagen and fibronectin. A significant improvement in the expression of type II collagen and aggrecan mRNA could be achieved. In addition, the monolayer cultures showed better synthesis of the extracellular matrices than alginate-bead cultures in SFM-P.

• KSBB Journal
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• 제8권1호
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• pp.17-22
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• 1993

$1.96{\times}10^{-5}$(IU/cell/hr) of specific scu-PA production rate was obtained from HEK cells in maintaining ca. $8{\times}10^{5}$(cells/ml) of maximum roll density at 10(ml/hr) of perfusion rate with recycling 20% serum free conditioned media. It can be compared to $4{\times}10^{6}$(cells/ml) of maximum cell density and $4.56{\times}10^{-4}$(IU/cell/hr) of specific production rate in cultivating cells with 1% serum containing medium. Thc conversion ratio of scu-PA to tc-UK increased up to 55% as the recycling ratio increased; however, recycling the used medium seemed to have least negative effect on cell growth. It also showed that the recycling process had definitive advantage of using serum free medium in perfusion cultivation of HEK cell line.

• KSBB Journal
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• 제8권1호
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• pp.23-27
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• 1993

• KSBB Journal
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• 제13권2호
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• pp.181-186
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• 1998

부착의존성 세포주인 Trichoplusia ni 의 유래의 BTI-TN5B1-4 (TN5) 곤충세포주를 이용하여 인간 혈소판생성축진인자인 재조합 인간 트롬보포이에틴(rhTPO)의 배양조건 최적화 연구를 수행하였다. 배양배지, 세포감염에 투입되는 재조합 베큘로바이러스와 숙주세포의 비율(MOI),세포감염시 세포밀도, 배지 회수시간 및 배양방법 등이 rhTPO 의 생산에 미치는 효과를 연구하여 60 mm dish로 정체 배양시 10 MOl 이상,$2\times10^6$ cells 의 세포밀도,바이러스 감염 후72 시간에서 rhTP0 의 최대 발현양 (약 12 mg/L)을 나타내었다. 배양 배지로서는 EXCELL FIVE 배지가 SF900II나Insect serum free media-1 Figure 5. Effect of growth phases on rhTPO production. TN5 cells were grown as suspension culture in 1 L spinner flask with 200 mL of SF900II serum free medium at 80 rpm. The cells were infected with AcBac404-2 at MOl of 1. Culture medium was collected at given time intervals and the expression level of rhTPO was analyzed by ELISA (A) or immunoblot analysis (B). Lanes 1 and 7; cell density of $0.6\times10^6$ cells/mL, lanes 2 and 8; cell density of $1.6\times10^6$ cells/mL, lanes 3 and 9; cell density of $2.0\times10^6$ cells/mL, lanes 4 and 10; cell density of $3.0\times10^6$ cells/mL, lane M; prestained molecular weight marker (Bio-Rad). Lanes 1, 2, 3, and 4; culture medium was collected at 48 hpi and lanes 7, 8, 9, and 10; culture medium was collected at 72 hpi. Figure 6. Effect of culture media on rhTPO production. TN5 cells grown with different culture media were infected with AcBac-404-2 at 10 MOL 10$\mu$L of culture medium was run on SDS-PAGE and Immunoblot analysis was performed. Lane ];TN5 cells cultured with SF900II serum free media(Gibco),and lane 3; TN5 cells cultured with EXPRESS FIVE serum free media (Gibco) 에 비해 더 증가된 발현양을 나타내었다. TN5 세포주를 0.2 L 규모 (1 L spinner flask)oJl에서 세포간의 응집현상 없이 부유배양에 적응,배양시킨 후 세포성장 시기에 따른 발현을 조사한 결과 1 MOI의 감염조건 하에서는 $0.6\times10^6$cell/mL의 early exponential시기의 세포밀도에서 72시간 배양하였을 대 최대 발현양을 나타내었다. 나타내었다.

• Biotechnology and Bioprocess Engineering:BBE
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• 제8권2호
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• pp.142-146
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• 2003

Three insect cell lines, Sf9, Sf21 and Tn5Bl-4, and four different kinds of serum free media (SFM), Sf 900 II, EX-CELL 420, EX-CELL 405 and Express Five, were used to compare the nutrient consumption, byproduct formation, production of recombinant protein and protease activity in suspension cultures. The Sf 900 II SFM was a ppropriate for the cell growth and protein production of the Sf9 and Sf21 cell lines. When the Tn5Bl-4 cell line was grown in the Express Five SFM, the specific growth rate was 1.6 fold higher than those of either the Sf9 or Sf21 cell lines. The glucose and glutamine consumption rates per cells, were 4 and 2.3 times higher than those of the Sf9 cell line, respectively. The overall yield coefficients of the lactate and ammoniumion were 2.8 and 1.5 times higher compared to those of the Sf9 cell line. respectively. The maximum specific ${\beta}$-galactosidase production rate was 4.5 fold that of the Sf9 cell line, a 3 times higher protease activity per cell.

• 한국축산식품학회지
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• 제42권5호
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• pp.775-799
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• 2022

The purpose of this review is to summarize studies that investigate blood and the main components of fetal bovine serum (FBS) in vertebrates, including major livestock, and review the current research on commercializing cultured meat. Detailed research on FBS is still lacking; however, some studies have shown that FBS consists of proteins, carbohydrates, growth factors, cytokines, fats, vitamins, minerals, hormones, non-protein nitrogen, and inorganic compounds. However, there are few studies on how the composition of FBS differs from blood or serum composition in adult animals, which is probably one of the main reasons for not successfully replacing FBS. Moreover, recent studies on the development of FBS replacers and serum-free media have shown that it is difficult to conclude whether FBS has been completely replaced or serum-free media have been developed successfully. Our review of the industrialization of cultured meat reveals that many basic studies on the development of cultured meat have been conducted, but it is assumed that the study to reduce or replace ingredients derived from fetuses such as FBS has not yet been actively developed. Therefore, developing inexpensive and edible media is necessary for the successful industrialization of cultured meat.

• KSBB Journal
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• 제11권3호
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• pp.329-339
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• 1996

The effects of ammonium ion on cell growth kinetics, monoclonal antibody productivity, and cell metabolism of hybridoma cells were investigated. The mouse-mouse hybridoma cell line VlIIH-8 producing mouse IgG2a was used as a model system. Ammonium ion showed an inhibitory effect on cell growth and monoclonal antibody production. New immobilized adsorbents were developed for the reduction of the inhibitory effect of ammonium ion. The ammonium ion selective zeolite, Phillipsite-Gismondine was entrapped in calcium alginate bead or in dialysis membrane and applied to the hybridoma cell culture system for the in situ removal of ammonium ion from culture media. The effects of ammonium the both serum supplemented and serum free media on the cell growth were studied by applying immobilized adsorbents of calcium alginate bead type. The results demonstrated a substantial enhancement in cell growth. Applying immobilized adsorbents of dialysis membrane type to serum supplemented media also resulted in the stimulation of cell growth, cell viability and monoclonal antibody production.

• 대한약리학회지
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• 제29권1호
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• pp.73-83
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• 1993

Steroid hormone의 하나인 ${\beta}-estradiol$이 serum-free medium에서 배양한 토끼의 신장 근위세뇨관 상피세포의 세포성장과 기능에 미치는 영향을 관찰한 바 다음과 같은 결과를 얻었다 1. Serum-free medium에서 토끼의 신장 근위세뇨관 상피세포는 ${\beta}-estradiol$ 1 nM의 농도에서 유의한 세포 성장 촉진 효과를 나타내었고, ${\beta}-estradiol$ 10 nM이상의 농도에서는 세포성장이 억제되었다. 2. ${\beta}-Estradiol$은 serum-free medium에서 성장촉진인자의 하나인 hydrocortisone을 뺀 조건하에서 세포 성장을 증가시키었다. 3. ${\beta}-Estradiol$은 hydrocortisone을 growth supplement로 넣어준 serum-free medium에서 토끼 신장의 근위세뇨관 상피세포의 성장을 촉진시키었다. 4. ${\beta}-Estradiol$은 Northern blot analysis에 의하여 확인한 alpha I (IV) collagen mRNA level에는 별다른 변화를 보이지 않으나, ${\beta}-actin$mRNA level은 증가되었다. 이상의 결과로 미루어 보아, serum-free 그리고 hormonally defined media에서 ${\beta}-estradiol이$ 토끼의 신장 근위세뇨관 상피세포의 성장 및 기능에 대하여 촉진적으로 작용하는 것은 cellular microfilament의 중요한 구성단백의 하나로 밝혀진 ${\beta}-actin$의 합성 증가에 기인하는 것으로 생각된다.

• Archives of Pharmacal Research
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• 제20권4호
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• pp.297-305
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• 1997

A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, $E_2$, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITCPNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.