• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.60-60
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• 2003

Embryonic stem (ES) cells have property of self-renewal and can differentiate into the cells of all three primary germ layers. Recently, many growth factors, alteration of culture condition and gene modifications have been used to differentiate mouse and human ES cells into specific cell types. This study was performed to evaluate the differentiation protocol for human ES cells to the endodermal lineage cells. Human ES cells (Miz-hESl ) were cultured on STO feeder layer mitotically inactivated with mitemycin C, and embryoid bodies (EBs) were formed by suspension culture. Differentiation protocol of EBs consisted of three steps: stage I, culture of EBs for 6 days with ITSFn medium; stage II, culture of stage I cells for 8 days with N2 medium ; stage III, culture of stage II cells for 22 days with N2 medium. mRNA levels of the endodermal lineage differentiation genes were analyzed by semi- quantitative RT-PCR. The Oct-4 expression, a marker of the pluripotent state, was detected in undifferentiated human ES cells but progressively decreased after EBs formation. Differentiating human ES cells expressed marker genes of endodermal differentiation and pancreatic islet cells. GATA4, a-fetoprotein, Glut-2, and Ngn3 were expressed in all stages. However, albumin and insulin were expressed in only stage III cells. The human ES cells can be differentiated into endodermal lineage cells by multiple step culture system using various supplements. We are developing the more effective protocols for guided differentiation of human ES cells.

• Journal of Periodontal and Implant Science
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• 제29권3호
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• pp.607-621
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• 1999

This study examined the human fibroblasts cell attachment to commercially pure titanium surface which had been instrumented by 3 types of periodontal instruments. Commercially pure titanium plates were uniformly scaled using plastic, stainless steel, titanium curette. these all experimental groups 65 undirectional strokes with the designated curettes. Alteration of the surfaces due to instrumentation was evaluated by Form Talysurf(R) and reported as Ra value(mean surface roughness). Then other experimental groups were immersed in a cell suspension of human gingival fibroblasts($1{\times}10^5$ cell/ml). After 3 days of culture, cell attachment and morphology was observed by SEM, and attached cell were counted by Hemocytometer. A significant difference in mean Ra value was observed for surface instrumented by metal curette compared to either control surface or surface instrumented by the plastic curette(P<0.01). No stastically significant difference was noted between control surface and those instrumented by the plastic curette. SEM observation showed that cell morphology and attachment to the commercially pure titanium plate was similar appearance on the all experimental groups. Experimental groups instrumented by titanium curette and stainless steel curette were more attached cell number than control group, but experimental group instrumented by plastic curette were similar with control groups(P<0.01). In summary, metal curette produced an significant alteration of the commercially pure titanium surface and more favorable surface topography for cell attachment. Otherwise plastic curette was insignificantly altered the commercially pure titanium surface(P<0.01).

• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1176-1184
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• 2013

Anti-hepatocyte growth factor (anti-HGF) monoclonal antibodies (mAbs) are potential therapeutics against various cancers. Screening for high-producer clones is a time-consuming and complex process and is a major hurdle in the development of therapeutic mAbs. Here, we describe an efficient approach that allows the selection of high-producer Chinese hamster ovary (CHO) cell lines producing the novel anti-HGF mAb SFN68, which was generated previously by immunizing HGF bound to its receptor c-Met. We selected an SFN68-producing parental cell line via transfection of the dihydrofolate reductase-deficient CHO cell line DG44, which was preadapted to serum-free suspension culture, with an SFN68-expression vector. Subsequent gene amplification via multiple passages of the parental cell line in a methotrexate-containing medium over 4 weeks, followed by clonal isolation, enabled us to isolate two cell lines, 2F7 and 2H4, with 3-fold higher specific productivity. We also screened 72 different media formulated with diverse feed and basal media to develop a suboptimized medium. In the established suboptimized medium, the highest anti-HGF mAb yields of the 2F7 and 2H4 clones were 842 and 861 mg/l, respectively, which were about 10.5-fold higher than that of the parental cell line in a non-optimized basal medium. The selected CHO cell lines secreting high titers of SFN68 would be useful for the production of sufficient amounts of antibodies for efficacy evaluation in preclinical and early clinical studies.

• 공업화학
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• 제31권1호
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• pp.13-18
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• 2020

실크의 섬유 고분자 단백질인 피브로인을 사용하여 산화철 나노입자가 내포된 테라그노시스가 가능한 마이크로캡슐을 제조하였다. 열중량 분석으로 산화철의 함량은 4.28%, 자력계로는 5.11%로 측정되었다. 산화철 마이크로캡슐이 첨가된 마우스 섬유아세포 3T3 배양액에서 얻어진 세포 현탁액은 자력에 반응하여 맑게 변하고, 세포는 자석 방향으로 응집하였다. 배양접시 상단에 올려둔 네오디뮴 자석은 세포를 배양액 표면 중심으로 세포를 끌어모았다. 배양액 표면에 모인 세포들은 응집하여 72 h 이후 장축의 길이가 2 mm인 비대칭 타원체인 세포 집합체를 형성하였다. 세포집합체의 바깥층에는 세포들이 상대적으로 크고 서로 모여 치밀한 조직을 형성하였으나, 중심부는 물질전달제한으로 세포의 사멸이 진행되는 것으로 관찰되었다. 바깥층에는 산화철 마이크로캡슐이 자력의 방향으로 체인처럼 일렬로 늘어선 현상도 관찰되었다. 마이크로CT를 이용하여 세포응집체 내부의 산화철이 고루 분포하지 않고 자력 방향으로 비대칭적으로 분포하고 있음을 보였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 춘계학술발표대회
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• pp.317-320
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• 2000

유전자 재조합된 Nicotiana tabacum을 이용하여 동결 보존 후의 세포 생장을 관찰하였다. 형질 전환된 Nicotiana tabacum의 동결 보존에서 회복배지에 계면 활성제와 산소 전달 물질을 첨가한 경우 첨가하지 않은 경우에 비해 세포 생장을 증대 시켰고 산소 전달 물질 보다 계면활성제의 영향이 크다는 결과를 관찰하였다.

• 한국식물생명공학회:학술대회논문집
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• 한국식물생명공학회 2005년도 춘계학술대회 및 국제심포지움 초록집
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• pp.182-182
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• 2005

• 대한미생물학회지
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• 제22권1호
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• pp.23-33
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• 1987

To detect lepotospiral strains which produce hemolysin and determine the optimal condition for assaying hemolysin, we screened reference strains and observed some property of hemolysin. Hemolysin activity was assayed with cell free culture liquids and erythrocyte suspension. The production of hemolysin by local strains isolated in Korea was assayed and compared with that of reference strains. The hemolysin was produced by 18 strains among 38 reference strains and 3 local strains isolated in Korea. The production of hemolysin began with growth of Leptospira cultured in EMJH medium and reached maximum at stationary phase. The optimum temperature for hemolytic activity was $37^{\circ}C$. At lower temperature the activity of hemolysin was decreased progressively. The hemolytic activity was completely inactivated after :30 minutes' exposure at $56^{\circ}C$. Hemolysis pattern was "hot-cold type" which showed increased hemolysis after cold incubation. The hemolysin was most active on sheep erythrocyte and less active on ox, goat, human and guinea pig erythrocyte with the decreasing order.

• KSBB Journal
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• 제11권1호
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• pp.107-114
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• 1996

당근세포(Daucus carota)로 부터 체세포배 생산성 증대를 위하여 체세포 배 현탁 배양액으로부터 추출한 세포외 분비물질을 이용하여 다음과 같은 효과를 측정하였다.: 분자량의 차이, 세포외 분비 물질 추출 시기, 주입시기, 주입농도에 따른 각종 체세포 배 생산성 증감 효과. 체세포배 생산량은 최대 1,500개/ml까지 얻었으며, 10kDa 이상 분자량이 포함된 세포외 분비물질을 초기에 주입시켰을 때 생산성이 특히 증가함을 보여 주었다.

• Journal of Plant Biotechnology
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• 제4권1호
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• pp.17-21
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• 2002

To establish an the mass production system of grape anthocyanin pigments through callus and cell suspension culture, the effects of various combinations of auxins and cytokinins on friable callus production were studied. for friable callus production, 2,4-D was superior to other regulators. IAA at 2 mg/L induced callus from stem and petiole while NAA resulted in rooting. Callus induction rate increased with the 2,4-D level, and stem segments were superior to leaf blade or petiole, showing nearly 100% with 1 and 2 mg/L 2,4-D from petiole and stem. Combined treatments of 2,4-D + kinetin and NAA + BA also yielded friable callus from stem segments. In treatments with 1 mg/L 2,4-D + 1 mg/L kinetin and 1 mg/L NAA + 1 mg/L BA, callus induction rate was nearly 100%. The combination effect of 2,4-D and BA on anthocyanin production was not significant.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 발생공학 국제심포지움 및 학술대회 발표자료집
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• pp.60-61
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• 2001

To produce reconstituted rabbit embryos with fetal fibroblasts, the present study was evaluated the efficiencies of the activation conditions as assessments of subsequent development and chromosome in the embryos. New Zealand White rabbits were used throughout the study. Fetal fibroblasts collected from 22-d of fetuses were cultured in DMEM＋10% FBS in 5% CO₂ in air. The culture was maintained for 10 passages. In every passage half of cell suspension were kept in frozen. (omitted)