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FGF-2와 덱사메타손이 지방기질세포의 증식 및 분화에 미치는 영향
이선영 ( Sun Young Lee ),손민정 ( Min Jung Shon ),김태호 ( Tae Ho Kim ),김석영 ( Suk Young Kim ),손영숙 ( Young Sook Son ),김신윤 ( Shin Yoon Kim ),박의균 ( Eui Kyun Park ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.4
Adipose tissue derived stromal cells (ATSCs) have been shown to be differentiated into several types of cells including osteoblasts. In the present study we have investigated the effects of fibroblast growth factor-2 (FGF- 2) and dexamethasone (Dex) on the proliferation and osteoblastic differentiation of ATSCs. A combination of FGF- 2 and Dex compared to untreated control stimulated proliferation of ATSCs up to 45% and 25% at day 5 and 7, respectively. Compared to FGF-2 alone, FGF-2 plus Dex further augmented proliferation of ATSCs by 15% (day 5) and 5% (day 7). In response to osteogenic stimulation, ATSCs expanded with FGF-2/Dex for 7 days were able to be differentiated into osteoblasts as revealed by increased calcium precipitation, and increased expression of osteocalcin, alkaline phosphatase and Runx2. Taken together, these results demonstrate that FGF-2 in combination with Dex stimulates proliferation of ATSCs and FGF-2/Dex-expanded ATSCs maintain a potential to be differentiated into osteoblasts.
동정맥루 기능이상 진단에 대한 이학적 검사의 정확도 : 정맥조영술 소견과의 비교 Comparison with Venographic Findings
최정란 ( Jung Ran Choi ),김영수 ( Young Soo Kim ),윤선애 ( Sun Ae Yoon ),원유동 ( Yoo Dong Won ),손영숙 ( Young Suk Son ),송우정 ( Woo Jung Song ),송호철 ( Ho Cheol Song ),김용수 ( Yong Soo Kim ),장윤식 ( Yoon Sik Chang ),방병기 ( 대한신장학회 2006 Kidney Research and Clinical Practice Vol.25 No.5
열처리한 자가혈청이 성인 골수간엽줄기세포의 증식과 조골세포 분화에 미치는 영향
박의균 ( Eui Kyun Park ),이선영 ( Sun Young Lee ),김태호 ( Tae Ho Kim ),정필훈 ( Phil Hoon Choung ),강길선 ( Gil Son Khang ),손영숙 ( Young Sook Son ),김석영 ( Suk Young Kim ),김신윤 ( Shin Yoon Kim ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.1
Tissue engineering using mesenchymal stem cells (MSCs) is a promising technology for treatment or regeneration of tissue defects. However, to put this technology into practical use, it is necessary to determine suitable biomaterials as well as to expand MSCs ex vivo. One major challenge of ex vivo expansion of MSCs is to avoid the use of fetal bovine serum (FBS), which is a fundamental culture supplement. In this study, we investigated effects of autologous serum on proliferation and osteoblastic differentiation of bone marrow stem cells (BMSCs) isolated from patients with skeletal diseases. We isolated BMSCs from 7 independent patients with avasular necrosis (AVN; 3 cases), osteoarthritis (OA; 2 cases), femoral head fracture (1 case) or hip dysplasia (1 case) and compared the effects of FBS, heat inactivated (HAS) and normal autologous serum (HA) on the proliferation of BMSCs. Proliferation analyses of BMSCs revealed that 5 out of 7 BMSCs cultured in 10 % HAS showed higher proliferation than cells cultured in 10 % FBS. Moreover, analysis of osteoblastic differentiation as assessed by alkaline phosphatase (ALP) staining, a marker for osteoblastic differentiation, showed that HAS, AS and FBS were capable of supporting osteoblastic differentiation of BMSCs at the similar level. Interestingly, mineralization was increased in BMSCs cultured in HAS (5 out of 6 BMSCs). These results demonstrate that HAS compared with conventional FBS has a better potential to stimulate proliferation and mineralization of BMSCs as well as to support their osteoblastic differentiation.
자가혈청하에서 FGF-2와 덱사메타손에 의한 골수중간엽줄기세포의 증식과 분화에 대한 효과
손민정 ( Min Jung Shon ),이선영 ( Sun Young Lee ),김태호 ( Tae Ho Kim ),유창국 ( Chang Kook You ),김석영 ( Suk Young Kim ),정필훈 ( Phil Hoon Choung ),손영숙 ( Young Sook Son ),박의균 ( Eui Kyun Park ),김신윤 ( Shin Yoon Kim ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.4
Previously we have shown that heat-inactivated autologous serum (HAS) has a potential to stimulate proliferation and ostoblastic differentiation of bone marrow stromal cells (BMSCs). In the present study we investigated whether stimulatory effects of HAS on proliferation and osteoblastic differentiation of BMSCs are further potentiated by fibroblast growth factor-2 (FGF-2) and dexamethasone (Dex). As expected, FGF-2 and Dex stimulated proliferation of BMSCs up to 17% at 5 days and 26% at 7 days of culture compared to HAS control. These results suggest that FGF-2 and Dex in the presence of HAS further stimulate proliferation of BMSCs. In order to examine whether BMSCs expanded with FGF-2, Dex and HAS harbor multipotency, the expanded cells were stimulated with either osteogenic or adipogenic cocktails. BMSCs expanded with FGF-2, Dex and HAS for 7 days were able to be differentiated into either osteoblasts or adipocytes. Taken together, these results demonstrate that FGF-2 and Dex in combination with HAS further stimulates proliferation of BMSCs and these expanded cells maintain potentials to be differentiated into either osteoblasts or adipocytes.
여러 종류의 성장인자와 덱사메타손이 골수간엽줄기세포의 증식에 미치는 효과
손민정 ( Min Jung Shon ),이선영 ( Sun Young Lee ),김태호 ( Tae Ho Kim ),김석영 ( Suk Young Kim ),강길선 ( Gil Son Khang ),손영숙 ( Young Sook Son ),신홍인 ( Hong In Shin ),김신윤 ( Shin Yoon Kim ),박의균 ( Eui Kyun Park ) 한국조직공학과 재생의학회 2006 조직공학과 재생의학 Vol.3 No.4
Bone marrow stromal cells(BMSCs) are pluripotent cells capable of differentiating into several types of cells and are thus an attractive cell source for connective tissue engineering. For use of BMSCs in there applications, ex vivo expansion is necessary to obtain sufficient numbers of cells. Various growth factors regulate the recruitment of progenitor cells and their proliferation and differentiation into matured cells. In this study we have investigated the effects of different growth factors and in combination with dexamethasone(Dex) on the proliferation of BMSCs. We found that at a low concentration of growth factors(1 ng/mL) including fibroblast growth factor(FGF)-2, epidermal growth factor(EGF), hepatocyte growth factor(HGF), transforming growth factor-ß(TGF-ß), vescular endothelial growth factor(VEGF) and platelet derived growth factor-BB(PDGF-BB), FGF-2 was most effective in stimulation of proliferation in BMSCs in vitro. In addition, when FGF-2, PDGF and EGF were combined with Dex, proliferation of BMSCs was further stimulated. Among the growth factors, it was shown that FGF-2 in combination with Dex showed most prominent effect on stimulation of BMSCs. These results demonstrate that a low concentration of FGF-2 incombination with Dex may be useful stimulants for ex vivo expansion of BMSCs.
탈미네랄화 골분(DBP)이 PLGA 지지체의 염증반응을 완화시킨다
최방실 ( Bang Sil Choi ),김순희 ( Soon Hee Kim ),윤선중 ( Sun Jung Yun ),하현정 ( Hyun Jung Ha ),김문석 ( Moon Suk Kim ),양영일 ( Young Il Yang ),손영숙 ( Young Sook Son ),강길선 ( Gil Son Khang ),이종문 ( John M. Rhee ),이해방 ( 한국조직공학과 재생의학회 2006 조직공학과 재생의학 Vol.3 No.3
We developed the demineralized bone particle(DBP) impregnated poly(lactide-co-glycolide)(PLGA) scaffolds(PLGA/DBP) to investigate the effect of adhesion, growth, viability and inflammatory reaction of the cells. PLGA/DBP scaffolds were prepared by solvent casting/salt leaching method and were characterized by porosimeter, and scanning electron microscopy. NIH/3T3 fibroblast cells were cultured in the PLGA/DBP scaffolds as well as film and MTT assay was used to assess the viability of cells. Also, human promyelocytic leukemia cells(HL-60) were cultured in the PLGA/DBP scaffolds. We observed IL-1ß? and TNF-α expression of HL-60 cells seeded in PLGA/DBP scaffold by RT-PCR. NIH/3T3 fibroblast cell seeded on PLGA/DBP film were more adhere and spread with increasing DBP content due to increasing hydrophilicity and bioactivity. The fluorescence intensity of the band of TNF-α and IL-1ß? gene was decreased with increasing the concentration of DBP. It seems that the DBP affected on the improvement of physicochemical properties of PLGA such as biocompatibility, wettability and inflammatory response.
하현정 ( Hyun Jung Ha ),김순희 ( Soon Hee Kim ),윤선중 ( Sun Jung Yoon ),고연경 ( Youn Kyung Ko ),이은경 ( Eun Kyung Lee ),손영숙 ( Young Sook Son ),김문석 ( Moon Suk Kim ),이종문 ( John M. Rhee ),강길선 ( Gil Son Khang ),이해방 ( 한국조직공학과 재생의학회 2006 조직공학과 재생의학 Vol.3 No.4
In order to fabricate tissue engineered biodisc, cells isolated from nucleus pulposus(NP) and annulus fibrosus(AF) have been characterized with sequential passages. NP and AF cells were separately by enzymatical digestion with 0.25wt% collagenase, respectively. Morphological changes were observed by phase contrast microscope and cell proliferation was counted by hemacytometer. To analyze the biosynthesis of glycosaminoglycan and gene expression of Type I and Type II collagen, safranin-O staining and RT-PCR have been carried out, respectively. Proliferation of NP and AF cells increased up to passage 3 and passage 6. Morphology of NP cells was maintained to passage 5 and it of AF cells was maintained to passage 4. In the result of safranin-O staining, NP and AF cells was stained positively. Type II collagen gene of NP cells was not expressed after passage 6 and expression of Type I collagen gene of AF cells was maintained up to passage 9. It can be expected that these results provide the important information for the application of tissue engineered biodisc.
홍원선,홍석일,박인철,손영숙,정훈용,양석균,김해련,민영일 울산대학교 의과대학 1996 울산의대학술지 Vol.5 No.1
cathepsin L은 lysosomal cysteine 단백분해효소로서 기저막(basement membrane)과 세포외기질(extracellular matrix)을 파괴하여 암세포의 침윤과 전이에 중요한 역할을 하는 물질로 알려져 있다. 이러한 cathepsin L에 대한 mRNA 발현도를 5개의 사람 위선암(gastric adenocarcinoma) 세포주와 5명의 위선암 환자에서 채취한 조직에서 방사능으로 표지된 cathepsin L특이 cDNA를 사용한 Northern blot법으로 측정하였다. 위암의 전이병소에서 수립한 세포주인 SNU-5, SNU-16, MKN-45와 Kato Ⅲ에서는 cathepsin L mRNA가 발현되었으나 원발병소에서 수립한 AGS 세포주에서는 mRNA의 발현이 관찰되지 않았다. 5명의 위암 환자에서는 원발병소, 전이가 확인된 임파절 및 암 근처 정상 위점막에서 각각 조직을 채취하여 cathepsin L mRNA의 발현을 측정하였다. 원발병소와 전이병소에서는 모두 cathepsin L mRNA가 발현되었으나 정상 위점막조직에서는 전예에서 mRNA 발현이 관찰되지 않았다. 한편 mRNA의 발현도는 1예에서는 전이병소가 원발병소에 비해 높았으나, 2예에서는 전이병소에서 발현도가 낮았으며, 나머지 2예에서는 원발병소와 전이병소 사이에 차이가 없어, 원발병소와 전이병소 사이에 mRNA의 발현도의 일관성 있는 경향은 관찰되지 않았다. 이상의 결과는 cathepsin L은 위암의 발생과 진행에 있어 암세포의 침윤과 전이를 촉진하는 것 이외에 또 다른 역할을 할 가능성을 시사하고 있다고 사료된다. Cathepsin L, a lysosomal cysteine protease, is known to play an important role in cancer invasion and metastasis by degrading the components of basement membrane and extracellular matrix. The mRNA expression of cathepsin L was determined by Northern blot analysis using a radiolabeled cDNA specific for cathepsin L in five human gastric adenocarcinoma cell lines and five surgical specimens of primary gastric adenocarcinomas, their metastatic lymph nodes and matched adjacent normal mucosae. The mRNA of cathepsin L was expressed in all of the four cell lines established from the metastatic sites, SNU-5, SNU-16, MKN-45 and Kato Ⅲ, while not detected in one cell line established from the primary site, AGS. The mRNA was expressed in all of the five primary and five metastatic cancer specimens tested, while it was not detected in all matched normal mucosae. The intensities of the mRNA expressions, however, did not show the consistent pattern between primary sites and metastatic lymph nodes. These results suggest that cathepsin L may have the other function in addition to facilitation of the invasion and metastasis during the development and progression of stomach cancer.