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SIS/PLGA 필름에서 SIS 함량이 신경계 세포의 부착과 증식에 미치는 영향
김초민 ( Cho Min Kim ),김순희 ( Soon Hee Kim ),송이슬 ( Yi Seul Song ),정수현 ( Su Hyun Jung ),김혜린 ( Hye Lin Kim ),오재민 ( Jae Min Oh ),박종학 ( Jong Hak Park ),이동원 ( Dong Won Lee ),이종문 ( John M Rhee ),강길선 ( Gil Son K 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4
Porcine small intestinal submucosa(SIS) derived from the submucosal layer of porcine intestine, both generate a minimal inflammatory response following implantation and have been approved by the FDA for many types of biomedical applications. Poly(lactic-co-glycolic acid)(PLGA) is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. Also, it can be easily fabricated into many shapes. In this study, we fabricated natural/synthetic biomaterial hybrid films using 0, 10, 20, 40 and 80 wt% of SIS. Schwann cells(SCs) and olfactory ensheathing cells(OECs) were seeded on SIS/PLGA film and confirmed the effects of adhesion and proliferation on SCs and OECs according to content of SIS. We confirmed that specific 10~20 wt% SIS in SIS/PLGA films was superior to adhesion and proliferation of SCs and OECs and content of SIS such as 40 wt% and 80 wt% could not help cell growth and proliferation.
슈반세포와 골수간엽줄기세포의 공배양이 골수간엽줄기세포의 분화와 증식에 미치는 영향(2)
김초민 ( Cho Min Kim ),김순희 ( Soon Hee Kim ),송이슬 ( Yi Seul Song ),정수현 ( Su Hyun Jung ),박종학 ( Jong Hak Park ),오재민 ( Jae Min Oh ),이동원 ( Dong Won Lee ),유일수 ( Il Sou Yoo ),이종문 ( John M Rhee ),강길선 ( Gil Son Kh 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4
Schwann cell(SC) play an important role as they are the source of different trophic substances and present a great neurotrophic activity. Bone marrow stromal cells(BMSCs) contain mesenchymal stem cell and progenitor cells. BMSCs can differentiate into various cell types, including muscle and brain cell. The aim of this study is to investigate the differentiation of BMSCs through co-culture system with SCs. Pure SCs and BMSCs were cultured as control group. Expression of GFAP, P75 and NF were determined by immunocytochemical staining and reverse transcriptase polymer chain reaction(RT-PCR) was conducted to confirm mRNA expression of NF and NSE in BMSCs co-cultured with SCs. Following co-culture time with SCs, BMSCs underwent morphological changes similar to those of cultured SCs and stained positively for GFAP, P75 and NF protein. Electrophoresis analysis revealed that the levels of NF and NSE on BMSC co-cultured with SCs related with other group maintained up to 21 days. Our results showed that BMSCs able to differentiate into SC-like cells using co-culture system in vitro. These findings could be helpful for the development of the cell-based therapeutic strategies for CNS repair.
김초민 ( Cho Min Kim ),김수미 ( Su Mi Kim ),김순희 ( Soon Hee Kim ),이일우 ( Il Woo Lee ),김문석 ( Moon Suk Kim ),이종문 ( John M. Rhee ),강길선 ( Gilson Khang ),이해방 ( Hai Bang Lee ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.1
We have investigated differentiation of bone marrow stromal cells(BMSCs) through co-culture system with Schwann cells(SCs). Expression of the S-100, Schwann cell markers, was determined by immunocytochemical staining and reverse transcriptase polymer chain reaction(RT-PCR). The morphology and growth in terms of differentiated BMSC was observed by invert microscope. Following co-culture time with SCs, BMSCs underwent morphological changes similar to those of cultured SCs and stained positively for S-100 protein as well as high proliferation aspect were detected. Electrophoresis analysis revealed that the levels of S-100 on BMSC co-cultured with SCs related with other group maintained up to 21 days. These findings support the assertion that BMSCs can be differentiated into cells that are Schwann cell-like in terms of phenotype using Schwann cell only.
PLGA/DBP지지체에 파종된 신경계세포가 골수간엽줄기세포의 분화와 증식에 미치는 영향
김수미 ( Su Mi Kim ),김순희 ( Soon Hee Kim ),김초민 ( Cho Min Kim ),오아영 ( A Young Oh ),김근아 ( Geun Ah Kim ),이일우 ( Il Woo Lee ),이종문 ( John M. Rhee ),강길선 ( Gilson Khang ),김문석 ( Moon Suk Kim ),이해방 ( Hai Bang Lee ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.4
Poly(lactic-co-glycolic acid)(PLGA) is a biodegradable synthetic polymer with acceptable mechanical strength and has well-controlled degradation rate. Demineralized bone particle(DBP) contains powerful bioactive molecules. Bone marrow mesenchymal stem cells(BMSCs) are important progenitor and supporting cells that have the intrinsic ability to self-renewal and differentiate into multiple functional cells. In this study, we fabricated natural/ synthetic hybrid PLGA scaffolds included 20% of DBP(PLGA/DBP). Schwann cells(SCs) and olfactory ensheathing cells(OECs) were seeded on PLGA/DBP scaffolds. We confirmed that effects of PLGA/DBP scaffolds seeded OEC and SC on proliferation and differentiation of BMSCs. PLGA/DBP scaffolds seeded OECs and SCs were superior to differentiation into neuronal cell proliferation of BMSCs. This phenomenon was expected due to cytokine secreted by OECs and SCs promote proliferation and differentiation of BMSCs.
다양한 함량의 DBP를 함유한 PLGA 지지체에서의 OEC의 증식 및 표현형 분석
김수미 ( Su Mi Kim ),김순희 ( Soon Hee Kim ),김초민 ( Cho Min Kim ),오아영 ( A Young Oh ),이일우 ( Il Woo Lee ),김문석 ( Moon Suk Kim ),이종문 ( John M. Rhee ),강길선 ( Gilson Khang ),이해방 ( Hai Bang Lee ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.4
Olfactory ensheathing cells(OEC) have been reported to migrate long distances and to guide axonal regeneration after spinal cord injury(SCI). Bone morphogenetic proteins(BMPs), members of the TGF-β family of signaling molecules, have numerous functions in nervous system development. In this study, Therefore, we investigated interaction of OEC with poly(lactide-co-glycolide)(PLGA)/demineralized bone particle(DBP) scaffold according to content of DBP. We manufactured PLGA/DBP scaffold by solvent casting/salt leaching method. Cell proliferation seeded into the scaffold were assayed MTT test and NF, S-100, P75 and BDNF were expressed in OEC cultured PLGA/DBP scaffold according to content of DBP. We examined the cell adhesion and proliferation by scanning electron microscopy observation. We confirmed that OEC adhesion and proliferation was increased on PLGA/ DBP scaffold of higher content of DBP. In these results, we recognized that PLGA/DBP scaffold provide the suitable environment for improvement of cell proliferation and formation of extra cellular matrix.
김수진 ( Soo Jin Kim ),조은혜 ( Eun Hye Jo ),김초민 ( Cho Min Kim ),홍희경 ( Hee Kyung Hong ),이은용 ( Eun Yong Lee ),김순희 ( Soon Hee Kim ),강영선 ( Young Sun Kang ),주천기 ( Choun Ki Joo ),이종문 ( John M Rhee ),강길선 ( Gil So 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4
Retinal pigment epithelial cells(RPE) form a monolayer on the outer portion of sensory retina and have important physiologic function that include solute transport, phagocytosis and digestion of membranes shed from photoreceptor outer segment and drug detoxication. Recently, transplantation of normal RPE has been proposed as a potential therapeutic modality in the surgical management of subretinal neovascularization in age related macular degeneration. In this study, to develop an improved isolation method, we compared results between physical method and chemical method and selected a modified cell isolation technique. High cell attachment, rapid proliferation and high purity of retinal pigment eptihelial cells showed in chemical method II.
PLGA/DBP 필름에서 DBP 함량에 따른 슈반씨세포와 후각초성세포의 부착과 성장에 미치는 영향
김수미 ( Su Mi Kim ),김순희 ( Soon Hee Kim ),김초민 ( Cho Min Kim ),오아영 ( A Young Oh ),김문석 ( Moon Suk Kim ),강길선 ( Gilson Khang ),이종문 ( John M. Rhee ),이해방 ( Hai Bang Lee ) 한국조직공학·재생의학회 2007 조직공학과 재생의학 Vol.4 No.2
Poly(lactic-co-glycolic acid)(PLGA) is a biodegradable synthetic polymer with acceptable mechanical strength and well-controlled degradation rate. Also, it can be easily fabricated into many shapes. Demineralized bone particle(DBP) contains powerful bioactive molecules. In this study, we fabricated natural/synthetic biomaterial hybrid films using 0, 10, 20, 40 and 80 % of DBP. Schwann cells(SCs) and olfactory ensheathing cells(OECs) were seeded on PLGA/DBP film and confirmed the effects of adhesion and proliferation on SCs and OECs according to content of DBP. PLGA/DBP composite films are easily fabricated and provided bioactive environment that enhance the cell adhesion and proliferation. In this study, we confirmed that specific 10~20% DBP in PLGA/DBP films was superior to adhesion and proliferation of SCs and OECs and content of DBP such as 40% and 80% could not help cell growth and proliferation. These results suggest that DBP, natural material, provide suitable surface to neural cells and proper content affect on culture condition to improve cell adhesion and proliferation. In addition, these information would contribute to design of 3-dimentional PLGA/DBP scaffold for tissue engineered spinal cord regeneration.
케라틴/PLGA 지지체가 골수간엽줄기세포의 신경분화에 미치는 영향
오아영 ( A Young Oh ),김순희 ( Soon Hee Kim ),김초민 ( Cho Min Kim ),정수현 ( Su Hyun Jung ),전나리 ( Na Ri Jeon ),홍현혜 ( Hyun Hye Hong ),( Sang Jin Lee ),( Mark Van Dyke ),( James J Yoo ),이동원 ( Dong Won Lee ),이종문 ( 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.1
Bone marrow stromal cells(BMSCs) exhibit multiple traits of a stem cell population, and they can expand many times in vitro and be induced to differentiate into multiple cell types. Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. We think keratins are useful as natural biomaterial. We examined the effect of Keratin/PLGA scaffold on the neural differentiation of rat BMSCs. We developed the keratin loaded poly(L-lactide-co-glycolide)(PLGA) scaffolds. Keratin/PLGA 0, 20 and 50 wt% scaffolds were prepared by solvent casting/salt leaching method. BMSCs were harvested from the femurs of adult female Fischer rat. These cells were seeded in prepared Keratin/PLGA scaffold and cultured in Medium 50 ml DMEM, 2 %DMSO, 200 ?M BHA, 25 ?M valproic acid, 10 ?M forskolin, 1 ?M hydro-cotisone, 5?g/ml insulin for 5 day. The effect of Keratin/PLGA scaffold on the neural differentiation of rat BMSCs were assessed in culture using the MTT assay and RT-PCR was conducted to confirm mRNA expression of NSE and Nf for neural marker. We confirmed that effect of Keratin/PLGA scaffold on the neural differentiation of rat BMSCs. According to our results, 20 wt% keratin/ PLGA scaffold have good cell compatibility and the expression of neural markers was higher. In conclusion, Keratin/ PLGA scaffold, on which neural differentiation of rat BMSCs was possible.