• International Journal of Industrial Entomology and Biomaterials
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• 제27권2호
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• pp.303-311
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• 2013

In this study, we compared the efficiency of osteoblast differentiation media (ODM) containing three distinct reagent combinations in osteoblastic differentiation of human bone marrow-derived mesenchymal stem cells (hBMSCs) in monolayer culture. In addition, we analyzed growth and differentiation of hBMSCs on silk scaffolds and examined the bone-forming activity of a nanofibrous silk scaffold in a tibia diaphysis defect model of a rat hind limb with intramedullary nailing. Although all three ODM increased alkaline phosphatase activity to a comparable extent, the ODM containing bone morphogenetic protein-2 (BMP-2) was found to be significantly less effective in promoting mineral deposition than the others. Growth of hBMSCs on sponge-form silk scaffolds was faster than on nanofibrous ones, while osteoblastic differentiation was apparent in the cells grown on either type of scaffold. By contrast, bone formation was observed only at the edge of the nanofibrous scaffold implanted in the tibia diaphysis defect, suggesting that use of the silk scaffold alone is not sufficient for the reconstitution of the long bone defect. Since silk scaffolds can support cell growth and differentiation in vitro, loading MSCs on scaffolds might be necessary to improve the bone-forming activity of the scaffold in the long bone defect model.

• Experimental and Molecular Medicine
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• 제50권11호
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• pp.8.1-8.14
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• 2018

We previously demonstrated that the direct transplantation of human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs) into the dentate gyrus ameliorated the neurological symptoms of Niemann-Pick type C1 (NPC1)-mutant mice. However, the clinical presentation of NPC1-mutant mice was not fully understood with a molecular mechanism. Here, we found 14,15-epoxyeicosatrienoic acid (14,15-EET), a cytochrome P450 (CYP) metabolite, from hUCB-MSCs and the cerebella of NPC1-mutant mice and investigated the functional consequence of this metabolite. Our screening of the CYP2J family indicated a dysregulation in the CYP system in a cerebellar-specific manner. Moreover, in Purkinje cells, CYP2J6 showed an elevated expression level compared to that of astrocytes, granule cells, and microglia. In this regard, we found that one CYP metabolite, 14,15-EET, acts as a key mediator in ameliorating cholesterol accumulation. In confirming this hypothesis, 14,15-EET treatment reduced the accumulation of cholesterol in human NPC1 patient-derived fibroblasts in vitro by suppressing cholesterol synthesis and ameliorating the impaired autophagic flux. We show that the reduced activity within the CYP system in the cerebellum could cause the neurological symptoms of NPC1 patients, as 14,15-EET treatment significantly rescued cholesterol accumulation and impaired autophagy. We also provide evidence that the intranasal administration of hUCB-MSCs is a highly promising alternative to traumatic surgical transplantation for NPC1 patients.

• Molecules and Cells
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• 제39권5호
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• pp.418-425
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• 2016

Resveratrol (RES) plays a critical role in the fate of cells and longevity of animals via activation of the sirtuins1 (SIRT1) gene. In the present study, we intend to investigate whether RES could promote the self-renewal and neural-lineage differentiation in human umbilical cord derived MSCs (hUC-MSCs) in vitro at concentrations ranging from 0.1 to $10{\mu}M$, and whether it exerts the effects by modulating the SIRT1 signaling. Herein, we demonstrated that RES at the concentrations of 0.1, 1 and $2.5{\mu}M$ could promote cell viability and proliferation, mitigate senescence and induce expression of SIRT1 and Proliferating Cell Nuclear Antigen (PCNA) while inhibit the expression of p53 and p16. However, the effects were reversed by 5 and $10{\mu}M$ of RES. Furthermore, RES could promote neural differentiation in a dose-dependent manner as evidenced by morphological changes and expression of neural markers (Nestin, ${\beta}III-tubulin$ and NSE), as well as pro-neural transcription factors Neurogenin (Ngn)1, Ngn2 and Mash1. Taken together, RES exerts a dosage-dependent effect on the self-renewal and neural differentiation of hUC-MSCs via SIRT1 signaling. The current study provides a new strategy to regulate the fate of hUC-MSCs and suggests a more favorable in vitro cell culture conditions for hUCMSCs-based therapies for some intractable neurological disorders.

• 한국발생생물학회지:발생과생식
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• 제14권4호
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• pp.233-241
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• 2010

사람의지방줄기세포는지방조직내에존재하는 줄기세포로 얻기 쉽고, 골수줄기세포와 유사한 특징을 가지고 있다. 그러나 지방을 추출하는 과정, 공여자의 나이, 체질량, 추출 부위에 따라 세포의 특성이 달라지며, 이질적인 세포군을 얻게 된다. 따라서 본 연구에서는 허벅지 지방에서 유래한 줄기세포 특성 분석 및 중배엽, 내배엽성 세포로의 분화능을 알아보았다. 허벅지 유래 줄기세포는 골수줄기세포와 유사한 섬유아세포와 유사한 모양을 보였으며, 체외에서 56.5번의 분열을 하였고, 약 $5{\times}10^{22}$개의 세포를 얻을 수 있었다. 이들은 SCF, Oct4, nanog, vimentin, CK18, FGF5, NCAM, Pax6, BMP4, HNF4a, nestin, GATA4, HLA-ABC, HLA-DR과 같은 유전자를 발현하였으며, Oct4, Thy-1, FSP, vWF, vimentin, desmin, CK18, CD54, CD4, CD106, CD31, a-SMA, HLA-ABC 등과 같은 단백질을 발현하였다. 또한 이들은 지방, 골, 연골 세포와 같은 중배엽성 세포로 분화하였고, 더욱이 인슐린 분비세포와 같은 내배엽성 세포로도 분화하였다. 결론적으로, 사람의 허벅지 유래 줄기세포는 골수 줄기세포와 유사하게 체외에서 증식이 가능하였으며, 유전자 및 단백질 발현 패턴을 가지고 있었으며, 다양한 세포로 분화 가능하였다. 이러한 결과로 미루어 보아 허벅지 지방유래 줄기세포는 골수 줄기세포를 대체할 수 있는 세포치료제의 재료가 될 수 있을 것으로 사료된다.

• Experimental and Molecular Medicine
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• 제50권12호
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• pp.12.1-12.14
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• 2018

Osteoporosis develops with high prevalence in both postmenopausal women and hypogonadal men. Osteoporosis results in significant morbidity, but no cure has been established. Mesenchymal stem cells (MSCs) critically contribute to bone homeostasis and possess potent immunomodulatory/anti-inflammatory capability. Here, we investigated the therapeutic efficacy of using an infusion of MSCs to treat sex hormone-deficient bone loss and its underlying mechanisms. In particular, we compared the impacts of MSC cytotherapy in the two genders with the aim of examining potential gender differences. Using the gonadectomy (GNX) model, we confirmed that the osteoporotic phenotypes were substantially consistent between female and male mice. Importantly, systemic MSC transplantation (MSCT) not only rescued trabecular bone loss in GNX mice but also restored cortical bone mass and bone quality. Unexpectedly, no differences were detected between the genders. Furthermore, MSCT demonstrated an equal efficiency in rectifying the bone remodeling balance in both genders of GNX animals, as proven by the comparable recovery of bone formation and parallel normalization of bone resorption. Mechanistically, using green fluorescent protein (GFP)-based cell-tracing, we demonstrated rapid engraftment but poor inhabitation of donor MSCs in the GNX recipient bone marrow of each gender. Alternatively, MSCT uniformly reduced the $CD3^+T$-cell population and suppressed the serum levels of inflammatory cytokines in reversing female and male GNX osteoporosis, which was attributed to the ability of the MSC to induce T-cell apoptosis. Immunosuppression in the microenvironment eventually led to functional recovery of endogenous MSCs, which resulted in restored osteogenesis and normalized behavior to modulate osteoclastogenesis. Collectively, these data revealed recipient sexually monomorphic responses to MSC therapy in gonadal steroid deficiency-induced osteoporosis via immunosuppression/anti-inflammation and resident stem cell recovery.

• 한국임상수의학회지
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• 제29권1호
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• pp.82-86
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• 2012

5년령 중성화된 수컷 치와와가 수개월간의 오른쪽 후지 파행 증상으로 내원하였다. 신체 검사에서 통증을 동반한 4등급의 슬개골 내측 탈구를 확인 하였고 오른쪽 왼쪽 후지 파행등급 2 와 0을 각각 확인 하였다. 일반 방사선 사진에서 골증식체와 연골하 병변을 확인 하였고 오른쪽과 왼쪽 후지 골관절증 점수 20 과 12를 각각 확인하였다. 슬개골 탈구 정복술과 자가지방 유래 줄기세포 치료를 보호자의 동의 하에 결정 하였다. $1{\times}10^6$ 개의 줄기세포를 PBS, 인공 괄절액과 함께 슬개골 탈구 정복술 후에 주입 하였다. 주입 후 19개월에 오른쪽 왼쪽 후지 파행등급 0 와 0, 골관절증 점수 19 와 13을 각각 확인 하였고, 5년후에 파행등급 0 와 0, 골관절증 점수 14 와 15를 각각 확인 하였다. 본 증례를 통해 자가지방 유래 줄기 세포치료를 받은 관절에서 골관절증이 감소 된 것을 확인 할 수 있었다.

• 한국임상수의학회지
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• 제28권1호
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• pp.122-127
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• 2011

후지마비와 심부통각의 소실을 보이는 5 년령의 수컷 Cocker Spaniel이 내원하였다. 신체검사, 신경검사, 방사선검사와 컴퓨터 단층촬영결과 심부통각의 소실을 동반한 요추 2번과 3번 사이의 디스크 탈출증으로 진단하였다. 편측 추궁 절제술을 시행하여 감압술과 탈출된 디스크 물질을 제거하였으며 $1{\times}10^6$ 개의 동종 지방유래 줄기세포를 $50{\mu}l$ 생리식염수에 희석하여 손상된 척수에 직접 주입하였다. 수술 10주 후, 양쪽 후지 모두에서 심부통각과 운동기능이 완전히 회복되었으며 세포이식과 관련된 부작용은 현재까지 발견되지 않았다. 본 증례를 통해 심각한 정도의 디스크질환을 가진 환축에서 감압술과 병행된 줄기세포의 이식방법은 좀더 나은 예후를 기대하기 위한 치료방법들 중 하나로 고려해볼 수 있을 것으로 생각된다.

• Childhood Kidney Diseases
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• 제14권1호
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• pp.32-41
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• 2010

목 적 : 사구체신염은 흔히 단백뇨를 보이며 특이 치료법이 없고, 만성 신부전으로 발전하는 경우가 많다. 몇몇 연구에서 중간엽 줄기세포(Mesenchymal stem cell, MSC)를 실험적 사구체신염에 투여하여 단백뇨가 호전된 것을 보고한 바 있으나, 이는 신염을 일으키는 약제와 중간엽 줄기세포를 함께 투여하거나 신장에 직접 투여한 것이었다. 본 연구에서는 실험적 신병증에서 단백뇨가 발현된 시점에서 정주 요법으로 MSC를 투여함으로써 MSC의 임상적인 적용 가능성을 탐색하였다. 방 법 : 실험용 생쥐에 Adriamycin을 투여하여 신병증(ADR-GN)을 유발한 후, 2주 후에 대량의 단백뇨를 확인하고 MSC를 생쥐 꼬리의 정맥에 주사하였다. MSC에 의한 질병 완화의 기전을 확인하기 위한 in vitro 실험으로 mixed lymphocyte culture(MLC)에 MSC를 투여하였을 때의 염증 관련 cytokine인 IFN-$\gamma$ and IL-10의 변화를 측정하였다. 결 과 : 실험용 생쥐에 ADR-GN를 유발하고 단백뇨가 보일 때 MSC를 정주한 군에서는 단백뇨의 소실이 더 먼저 관찰되었다. 또한 MSC를 투여받은 군에서의 생존률이 더 나은 경향이 관찰되었다. MLC 에 MSC를 투여하였을 때, 염증을 유발하는 cytokine인 IFN-$\gamma$ 는 감소하고 염증을 억제하는 cytokine인 IL-10는 증가하였다. 결 론 : 이 연구는 이전의 보고들에서 관찰되었던 사구체신염에서의 MSC의 질병완화 효과가 좀더 임상적으로 적용 가능한 방법으로 투여된 경우, 즉 단백뇨가 있을 때 정주 요법으로 투여한 경우에도 관찰됨을 확인하였다. 이러한 효과의 기전과 임상적용에 요구되는 안전성 등에 대한 확인을 위해서는 추가 연구가 필요하겠다.

• Molecules and Cells
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• 제40권6호
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• pp.386-392
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• 2017

Periodontal ligament stem cells (PDLSCs) are multipotent stem cells derived from periodontium and have mesenchymal stem cell (MSC)-like characteristics. Recently, the perivascular region was recognized as the developmental origin of MSCs, which suggests the in vivo angiogenic potential of PDLSCs. In this study, we investigated whether PDLSCs could be a potential source of perivascular cells, which could contribute to in vivo angiogenesis. PDLSCs exhibited typical MSC-like characteristics such as the expression pattern of surface markers (CD29, CD44, CD73, and CD105) and differentiation potentials (osteogenic and adipogenic differentiation). Moreover, PDLSCs expressed perivascular cell markers such as NG2, ${\alpha}-smooth$ muscle actin, platelet-derived growth factor receptor ${\beta}$, and CD146. We conducted an in vivo Matrigel plug assay to confirm the in vivo angiogenic potential of PDLSCs. We could not observe significant vessel-like structures with PDLSCs alone or human umbilical vein endothelial cells (HUVECs) alone at day 7 after injection. However, when PDLSCs and HUVECs were co-injected, there were vessel-like structures containing red blood cells in the lumens, which suggested that anastomosis occurred between newly formed vessels and host circulatory system. To block the $SDF-1{\alpha}$ and CXCR4 axis between PDLSCs and HUVECs, AMD3100, a CXCR4 antagonist, was added into the Matrigel plug. After day 3 and day 7 after injection, there were no significant vessel-like structures. In conclusion, we demonstrated the perivascular characteristics of PDLSCs and their contribution to in vivo angiogenesis, which might imply potential application of PDLSCs into the neovascularization of tissue engineering and vascular diseases.

• 한국수정란이식학회:학술대회논문집
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• 한국수정란이식학회 2006년도 The 6th Intermational Symposium on Developmental Biotechnology
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• pp.15-15
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• 2006