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복합적인 기계적 자극이 중간엽 줄기세포의 골 분화에 미치는 영향
강윤경 ( Yun Gyeong Kang ),전강진 ( Kang Jin Jeon ),박소희 ( So Hee Park ),신지원 ( Ji Won Shin ),김선연 ( Seon Yeon Kim ),현진숙 ( Jin Sook Hyun ),신정욱 ( Jung Woog Shin ) 한국조직공학·재생의학회 2012 조직공학과 재생의학 Vol.9 No.2s
In this study we investigated the combinational stimulation effects of uniaxial tension and intermittent hydrostatic pressure (IHP) on the differentiation of mesenchymal stem cells (MSCs) into osteoblast without any biochemical agents. For this, we designed a novel bioreactor which can generate not only tensile stimulation and IHP but also combinational stimulation of tension and IHP. We classified experimental groups based on the types of stimulation as follows: (1) NS: no stimulation as a control; (2) IHP: IHP stimulation only (magnitude: 0.1 MPa, intermittent pattern of hydrostatic pressure: 2 min/15 min (on/off), duration: 3 hr/day, for 2 days); (3) TS: tensile stimulation only (magnitude: 3%, frequency: 0.5 Hz, 48 hr in a row); (4) CS: combinational stimulation of (2) and (3). We conducted various biological analyses; DNA contents, ALP activity, Live & Dead staining, F-actin staining, von Kossa staining, and ALP staining. As results, the groups applied mechanical stimulation (IHP, TS, and CS group) showed higher proliferation rate than the group applied no stimulation (NS) at day 8. And amounts of ALP normalized by DNA contents in the groups of TS and IHP were higher than those in CS group at day 5 and day 8, respectively. The highest expressions of ALP and von Kossa staining were observed in IHP group. The expressions of ALP and von Kossa staining in CS were negligible compared with IHP. From this study, we concluded that the single type of mechanical stimulation (IHP or tension) has positive effects but the effect of combinational mechanical stimulation was imperceptible on osteogenesis of MSCs.
다양한 간헐적 정수압 유형이 중간엽 줄기세포의 골 분화에 미치는 영향
강윤경 ( Yun Gyeong Kang ),( Garcia M V ),( Marquez J C ),박소희 ( So Hee Park ),오민재 ( Min Jae Oh ),김영미 ( Young Mi Kim ),신정욱 ( Jung Woog Shin ) 한국조직공학과 재생의학회 2014 조직공학과 재생의학 Vol.11 No.1s
This study investigated the effects of intemittent hydrostatic pressure (IHP) patterns on the responses of mesenchymal stem cells (MSCs) such as osteogenic differentiation, proliferation and senescence. For these experimental groups were set based on IHP patterns: (1) C_BM: control group with no stimulation in basal media; (2) C_OM: control group with no stimulation in osteogenic media (OM); (3) S_2H/5M: longer pressurizing (2hours) and shorter resting (5minutes) time in OM; (4) S_1H/1H: equal pressurizing and resting time (1hour) in OM; (5) S_2M/15M: shorter pressurizing (2minutes) and longer resting (15minutes) time in OM. The magnitude of IHP was 0.15 MPa. IHP was applied to corresponding groups for 4 hours a day from 3 days starting at 48 hours after seeding. Examination of DNA contents, ALP activity and its staining, quantitative real-time PCR, and β-galactosidase staining were performed. ALP amount normalized by corresponding DNA content in S_2H/5M, C_BM was significantly lower than that of the other group at day 5, which was more observable even at day 7 while S_2H/5M significantly had lower ALP count than other groups at day 5 and day 7. Other groups (S_1H/1H and S_2M/15M) showed significantly higher ALP amounts indicating the positive effect on osteogenic differentiation. Other markers indicating the degrees of differentiation showed comparable results. Based on β-galactosidase staining, it appeared that mechanical stimuli did not affect cell senescence significantly. From this study, we concluded that engagement of IHP has a potential of controlling osteogenic differentiation depending on its pattern: it can promote or suppress differentiation.
김태은 ( Kim Tae Eun ),강윤경 ( Kang Yun Gyeong ),정명철 ( Jeong Myeong Cheol ),노지현 ( No Ji Hyeon ),심의섭 ( Sim Ui Seob ),김용봉 ( Kim Yong Bong ) 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.4
The immature teratoma is the 3rd most common germ cell tumor and represents 10-20% of all ovarian malignancies occurring in the first two decades of life. The initial clinical manifestation is abdominal/pelvic mass with pain. This tumor is usually unilate
공배양 및 물리적 환경하에서의 중간엽 줄기세포의 혈관 관련 세포로의 분화 경향
박소희 ( So Hee Park ),신지원 ( Ji Won Shin ),김수향 ( Su Hyang Kim ),강윤경 ( Yun Gyeong Kang ),전강진 ( Kang Jin Jeon ),김선연 ( Seon Yeon Kim ),현진숙 ( Jin Sook Hyun ),신정욱 ( Jung Woog Shin ) 한국조직공학·재생의학회 2013 조직공학과 재생의학 Vol.10 No.1s
How to control the differentiation of mesenchymal stem cells (MSCs) into vascular lineage cells have still left much unclear even with numerous studies. Most in vitro studies on the differentiation of stem cells have utilized biochemical reagents. However, recent studies show the potentials of physical and co-culturing environments in control of differentiation. Therefore, we first factored out the in vivo conditions into biochemical reagents, co-culture, and physical stimulation. Then combinational effects of those factors on the differentiation of MSCs into vascular lineage cells were studied. For this EGM (R) -2, co-culture, and shear stress were adopted to meet biochemical, neighboring cells, and physical condition, respectively. A total 4 experimental groups were set depending on co-culture and shear stress. Cell density was 1×104cells/cm2 at all group. For the co-cultures, MSCs were mixed with human umbilical vein endothelial cells (HUVECs) in the ratio of 1:2. The 12 dyne/cm2 of shear stress was engaged: 4 hr/ day for 2 days followed by 48 hr after seeding. For the analyses, the immunofluorescent staining and PCR was performed to evaluate the differentiation of MSCs into vascular lineage cells. The co-culture group without shear stress showed the ECs-related markers even on the first day of the culture. And this was more observable on day 4. However, differentiation of MSCs into smooth muscle cells was found dominant when shear stress was engaged even MSCs were cultured with HUVECs and EGM (R) -2. From this study we confirmed that physical stimulation (12 dyne/ cm2 in our study) plays an important role in controlling differentiation of MSCs into vascular lineage cell types when MSCs were co-cultured with ECs.
물리적 인장 자극에 의한 줄기세포의 분화에 동반되는 미토콘드리아의 특성 변화에 관한 고찰
신지원 ( Ji Won Shin ),김혜림 ( Hye Lim Kim ),강윤경 ( Yun Gyeong Kang ),박소희 ( So Hee Park ),김영미 ( Young Mi Kim ),신정욱 ( Jung Woog Shin ) 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.1s
Recent studies have confirmed that the direction of stem cell differentiation can be modulated by physical stimuli even without biochemical growth factors. This implies stem cells attribute their differentiation to mechanical signaling directly or indirectly. However, the role or changes in mitochondria, one of the major energy sources in a cell, during differentiation have not been investigated. This study applied mechanical stretching of two different magnitudes (3% and 10%) to human mesenchymal stem cells without any relevant growth factors. We confirmed differentiation of stem cells into two different directions: osteogenic- and smooth muscle-like cells. Along with these results we found the expression of NOX4 was increased with stretching magnitude. As the expression of NOX4 is closely related to mitochondrial biogenesis and morphology various markers were also investigated. The expressions of the markers in relation to biogenesis (PGC-1alpha, TFA) and fusion (MFN1, MFN2, OPA1) were increased with stretching magnitude. However, the expression of Drp1 was observable with stretching of 3%. This study showed that the magnitude of physical stimulation affects the expression of NOX4, which has been known to be closely related to the biogenesis and morphological changes of mitochondria and stem cell differentiation. Therefore, further study of NOX4 due to physical stimulation for the control of stem cell differentiation accompanied by mitochondria functions is worth to be performed.
혼합배지와 지지체 특성에 의한 줄기세포 분화제어: 골연골 조직을 중심으로
한정윤 ( Jeong Yoon Han ),신지원 ( Ji Won Shin ),박소희 ( So Hee Park ),김동화 ( Dong Hwa Kim ),김수향 ( Su Hyang Kim ),강윤경 ( Yun Gyeong Kang ),신정욱 ( Jung Woog Shin ) 한국조직공학과 재생의학회 2011 조직공학과 재생의학 Vol.8 No.2s
This article investigated the potential of substrate characteristics for the control of differentiation of stem cells. For this, two distinctive materials, alginate and HA (hydroxiapatite) contained PCL, were selected. Utilizing rapid prototype (RP) technique we fabricated each type of scaffold. Mesenchymal stem cells (MSCs) derived from New Zealand White rabbits were suspended into alginate solution and the solution was plotted utilizing RP technique. The cocktail media which contain chondrogenic and osteogenic growth factors were used. The basal media were also used for comparison. During 14 days of experiments the samples were harvested four times. The results showed that MSCs tended to be differentiated into osteo- and chondrocyte-like cells when they were on PCL and in alginate scaffold, respectively even both of the two types of scaffolds were in basal media. These findings were more observable when the cocktail media were used. From this research we can stipulate that the characteristics of substrate materials should be considered along with biochemical reagents in the control of differentiation of stem cells. It is also recommended that the biomechanical factors, such as mechanical stimuli, should be considered in view of bio-mimetic environments.
PDMS 표면형상과 간헐적 정수압이 중간엽 줄기세포의 분화 및 증식에 미치는 영향
정재영 ( Jae Young Jeong ),김동화 ( Dong Hwa Kim ),박소희 ( So Hee Park ),신지원 ( Ji Won Shin ),한정윤 ( Jeong Yoon Han ),강윤경 ( Yun Gyeong Kang ),신정욱 ( Jung Woog Shin ) 한국조직공학과 재생의학회 2010 조직공학과 재생의학 Vol.7 No.3
We investigated the effects of PDMS surface patterns on differentiation and proliferation of MSCs along with intermittent hydrostatic pressure (IHP). Also the effect of growth factor was evaluated. For this, we fabricated PDMS substrates with or without patterns. The patterned substrates were made into two types: 5×5×3 μm 3 and 20×20×3 μm 3. The IHP (0.2 MPa, 2 hrs/day) was engaged on the all experimental groups for early 7 days starting 24 hrs after seeding the cells. To examine the cellular responses up to 21 days, we conducted various biological tests: DNA contents, ALP activity and GAG assay, RT-PCR for aggrecan, type II collagen, osteopontin, osteocalcin. The proliferation rates were higher in patterned groups without growth factor. The group with 20 μm pattern showed higher proliferation rate than group with 5 μm pattern. Amounts of GAG normalized by DNA contents in patterned groups were higher than those of non-patterned groups during the experiment periods and were remarkably higher in the groups with growth factor. ALP expressions were so small to be negligible even they tended to be decreased in all groups. activities tended to decrease in all groups. The expressions of chondrogenesis related genes (aggrecan, type II collagen) were observable in patterned groups. Osteopontin, the early marker of osteogenesis, was observable only in the early stage of experiment. Negligible expression of osteocalcin was detected in all groups. From this study, we concluded that patterned surface and IHP have positive effects on proliferation of MSCs and tended to enhance chondrogenesis rather than osteogenesis.