• 대한임상검사과학회지
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• 제50권4호
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• pp.391-398
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• 2018

중간엽줄기세포는 항염증능, 면역조절능 뿐만 아니라 다계통으로의 분화능 때문에 난치성 환자 치료를 위한 매력적인 대안적 치료방법으로 알려져 왔다. 지금까지 중간엽줄기세포의 이식 치료법은 면역질환, 심혈관질환, 암, 간질환 및 뇌졸중을 비롯한 다양한 질병의 전임상 및 임상적용에 긍정적인 결과를 가져왔다. 여러 연구들에 의하면, 중간엽줄기세포를 이용한 치료는 손상된 세포나 조직에 중간엽줄기세포가 이동하여 직접 세포를 대체하거나 분화시키는 작용이 아니라 중간엽줄기세포에서 분비하는 여러 인자들 즉, 주변분비 효과(paracrine effect)에 의한 것으로 확인되고 있다. 최근에 중간엽줄기세포 유래 엑소좀은 핵산, 단백질, 지질 등을 손상된 세포나 조직의 국소 미세환경으로 전달함으로써 세포간 상호작용을 통해 조직재생을 중재할 수 있는 중요한 역할을 하는 것으로 알려졌다. 엑소좀의 이용은 세포이식으로부터 발생할 수 있는 종양형성과 같은 다양한 위험성을 피할 수 있으므로 줄기세포 기반 치료 적용에 유용성이 매우 높다. 이러한 이유에서 중간엽줄기세포 유래 엑소좀은 재생의학 및 조직공학에서 안전하고 효율적인 치료적 도구(tool)가 될 수 있다. 여기에서 우리는 치료제로서의 중간엽줄기세포 유래 엑소좀의 정의와 역할에 대한 최신 지견과 함께 포괄적인 이해를 제공하고자 한다.

• 대한약침학회지
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• 제26권4호
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• pp.357-365
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• 2023

Objectives: Since stroke is a serious health issue, novel therapeutic strategies are required. In a mouse model of ischemic stroke, this study analyzed the potential of electroacupuncture (EA) and tenuigenin (TE) to improve the efficacy of human mesenchymal stem cell (hMSC) transplantation. Methods: Middle cerebral artery occlusion (MCAO) with reperfusion was used to generate ischemic stroke. Forty-eight male C57BL/6 mice were randomly divided into five groups: control, MCAO-operated, MCAO-EA, MCAO-TE, or MCAO + EA + TE. Subsequently, hMSCs were transplanted into the ischemic region and EA, TE, or the combination was administered. Behavior assessments and immunohistochemistry were conducted to evaluate motor and cognitive recovery and hMSCs survival, migration, and differentiation. Results: The combined treatment of EA and TE exhibited enhanced hMSCs survival, migration and differentiation into neural cell lineages while suppressing astrocyte formation. Immunohistochemistry demonstrated increased neurogenesis through hMSCs transplantation in the ischemic brain. Immediate behavioral improvements were not significantly different between groups, but there was a gradual recovery in motor and cognitive function over time. Conclusion: These findings highlight the potential of EA and TE co-treatment as a therapeutic strategy for ischemic stroke, opening avenues for further research to optimize treatment protocols and elucidate underlying mechanisms.

• International Journal of Stem Cells
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• 제16권3호
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• pp.251-259
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• 2023

Mesenchymal stromal cells (MSCs) have attracted scientific and medical interest due to their self-renewing properties, pluripotency, and paracrine function. However, one of the main limitations to the clinical application of MSCs is their loss of efficacy after transplantation in vivo. Various bioengineering technologies to provide stem cell niche-like conditions have the potential to overcome this limitation. Here, focusing on the stem cell niche microenvironment, studies to maximize the immunomodulatory potential of MSCs by controlling biomechanical stimuli, including shear stress, hydrostatic pressure, stretch, and biophysical cues, such as extracellular matrix mimetic substrates, are discussed. The application of biomechanical forces or biophysical cues to the stem cell microenvironment will be beneficial for enhancing the immunomodulatory function of MSCs during cultivation and overcoming the current limitations of MSC therapy.

• 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.

• Biomolecules & Therapeutics
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• 제23권6호
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• pp.517-524
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• 2015

Human mesenchymal stem cells (MSCs) have been used in cell-based therapy to promote revascularization after peripheral or myocardial ischemia. High levels of reactive oxygen species (ROS) are involved in the senescence and apoptosis of MSCs, causing defective neovascularization. Here, we examined the effect of the natural antioxidant lycopene on oxidative stress-induced apoptosis in MSCs. Although $H_2O_2$ ($200{\mu}M$) increased intracellular ROS levels in human MSCs, lycopene ($10{\mu}M$) pretreatment suppressed $H_2O_2$-induced ROS generation and increased survival. $H_2O_2$-induced ROS increased the levels of phosphorylated p38 mitogen activated protein kinase (MAPK), Jun-N-terminal kinase (JNK), ataxia telangiectasia mutated (ATM), and p53, which were inhibited by lycopene pretreatment. Furthermore, lycopene pretreatment decreased the expression of cleaved poly (ADP ribose) polymerase-1 (PARP-1) and caspase-3 and increased the expression of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax), which were induced by $H_2O_2$ treatment. Moreover, lycopene significantly increased manganese superoxide dismutase (MnSOD) expression and decreased cellular ROS levels via the PI3K-Akt pathway. Our findings show that lycopene pretreatment prevents ischemic injury by suppressing apoptosis-associated signal pathway and enhancing anti-oxidant protein, suggesting that lycopene could be developed as a beneficial broad-spectrum agent for the successful MSC transplantation in ischemic diseases.

• Reproductive and Developmental Biology
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• 제36권4호
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• pp.283-290
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• 2012

Skin serves as an easily accessible source of multipotent stem cells with potential for cellular therapies. In pigs, stem cells from skin tissues of fetal and adult origins have been demonstrated as either floating spheres (cell aggregates) or adherent spindle-shaped mesenchymal stem cell (MSC)-like cells depending on culture conditions. The cells isolated from the epidermis and dermis of porcine skin showed plastic adherent growth in the presence of serum and positively expressed a range of surface and intracellular markers that are considered to be specific for MSCs. The properties of primitive stem cells have been observed with the expression of alkaline phosphatase and markers related to pluripotency. Further, studies have shown the ability of skin-derived MSCs to differentiate in vitro along mesodermal, neuronal and germ-line lineages. Moreover, preclinical studies have also been performed to assess their in vivo potential, and the findings appear to be effective in tissue regeneration at the defected site after transplantation. The present review describes the recent progress on the biological features of porcine skin-derived MSCs as adherent cells, and summarizes their potential in advancing stem cell based therapies.

• 생명과학회지
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• 제17권5호
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• pp.678-686
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• 2007

인간중간엽줄기세포는(Human mesenchymal stem cells, hMSC) 골수, 지방, 피부, 근육, 혈액에 존재하며, 뼈, 연골, 지방, 근육, 신경세포로 분화가능성이 보고되어 손상조직의 재생을 위한 재료로서뿐만 아니라 유전자치료의 매개체로 이용될 수 있는 가능성이 제안되고 있다. 인간중간엽줄기세포의 적절한 배양조건에는 소 태아혈청(fetal bovine serum, FBS)이 요구되어지므로 세포치료에는 소 태아혈청이 다수 포함되어 있을 것이며 세포배양 배지 유래 소 태아혈청의 단백질에 의한 면역거부반응이 우려된다. 이미 앞선 연구에서 자가혈청 하에서 인체지방줄기세포 분리와 계속적인 세포배양을 실시하였을 때 인체지방줄기세포의 증식능력과 다 분화 능이 유지되며 면역결핍 생쥐에 골수의 말초혈액에서 유래된 CD34세포 이식 시 안착 능을 촉진함을 보였다. 본 연구에서 인체지방줄기세포가 인체혈청 하에서 배양되었을 때 소 태아혈청 하에서 배양할 때 발현하는 표면항원을 유지함을 확인했으며 microarray를 사용하여 유전자 발현을 비교했다. 유 세포 분석을 통하여 인체혈청 하에서 계속적으로 배양된 인체유래지방줄기세포에서 HLA-DR, CD117, CD29 와 CD44 의 발현이 소 태아혈청 하에서 배양했을 때와 비슷함을 밝혔다. 그러나 인체혈청 하에서 배양된 인체지방줄기세포의 유전자 발현형태와 소 태아혈청 하에서 배양된 세포의 유전자 발현형태 간에는 상당한 차이를 보였다. 그러므로 본 연구는 인체혈청 하에서 배양된 인체지방기질줄기세포가 임상적용을 위한 선행 데이터로써 직접적인 추정을 하기 위해서는 인체지방기질줄기세포 이식연구에 in vivo 동물실험연구가 수행되어져야 함을 제시하고 있다.

• Molecules and Cells
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• 제40권2호
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• pp.133-142
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• 2017

Previous studies have shown that bone marrow mesenchymal stromal cell (MSC) transplantation significantly improves the recovery of neurological function in a rat model of intracerebral hemorrhage. Potential repair mechanisms involve anti-inflammation, anti-apoptosis and angiogenesis. However, few studies have focused on the effects of MSCs on inducible nitric oxide synthase (iNOS) expression and subsequent peroxynitrite formation after hypertensive intracerebral hemorrhage (HICH). In this study, MSCs were transplanted intracerebrally into rats 6 hours after HICH. The modified neurological severity score and the modified limb placing test were used to measure behavioral outcomes. Blood-brain barrier disruption and neuronal loss were measured by zonula occludens-1 (ZO-1) and neuronal nucleus (NeuN) expression, respectively. Concomitant edema formation was evaluated by H&E staining and brain water content. The effect of MSCs treatment on neuroinflammation was analyzed by immunohistochemical analysis or polymerase chain reaction of CD68, Iba1, iNOS expression and subsequent peroxynitrite formation, and by an enzyme-linked immunosorbent assay of pro-inflammatory factors (IL-$1{\beta}$ and TNF-${\alpha}$). The MSCs-treated HICH group showed better performance on behavioral scores and lower brain water content compared to controls. Moreover, the MSC injection increased NeuN and ZO-1 expression measured by immunochemistry/immunofluorescence. Furthermore, MSCs reduced not only levels of CD68, Iba1 and pro-inflammatory factors, but it also inhibited iNOS expression and peroxynitrite formation in perihematomal regions. The results suggest that intracerebral administration of MSCs accelerates neurological function recovery in HICH rats. This may result from the ability of MSCs to suppress inflammation, at least in part, by inhibiting iNOS expression and subsequent peroxynitrite formation.

• Molecules and Cells
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• 제41권12호
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• pp.1016-1023
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• 2018

Regenerative orthopedics needs significant devices to transplant human stem cells into damaged tissue and encourage automatic growth into replacements suitable for the human skeleton. Soft biomaterials have similarities in mechanical, structural and architectural properties to natural extracellular matrix (ECM), but often lack essential ECM molecules and signals. Here we engineer mineralized polysaccharide beads to transform MSCs into osteogenic cells and osteoid tissue for transplantation. Bone morphogenic proteins (BMP-2) and indispensable ECM proteins both directed differentiation inside alginate beads. Laminin and collagen IV basement membrane matrix proteins fixed and organized MSCs onto the alginate matrix, and BMP-2 drove differentiation, osteoid tissue self-assembly, and small-scale mineralization. Augmentation of alginate is necessary, and we showed that a few rationally selected small proteins from the basement membrane (BM) compartment of the ECM were sufficient to up-regulate cell expression of Runx-2 and osteocalcin for osteoid formation, resulting in Alizarin red-positive mineral nodules. More significantly, nested BMP-2 and BM beads added to a non-union skull defect, self-generated osteoid expressing osteopontin (OPN) and osteocalcin (OCN) in a chain along the defect, at only four weeks, establishing a framework for complete regeneration expected in 6 and 12 weeks. Alginate beads are beneficial surgical devices for transplanting therapeutic cells in programmed (by the ECM components and alginate-chitosan properties) reaction environments ideal for promoting bone tissue.

• Biomolecules & Therapeutics
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• 제28권1호
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• pp.34-44
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• 2020

Mesenchymal stem cells (MSCs) have been proposed as an alternative therapy to be applied into several pathologies of the nervous system. These cells can be obtained from adipose tissue, umbilical cord blood and bone marrow, among other tissues, and have remarkable therapeutic properties. MSCs can be isolated with high yield, which adds to their ability to differentiate into non-mesodermal cell types including neuronal lineage both in vivo and in vitro. They are able to restore damaged neural tissue, thus being suitable for the treatment of neural injuries, and possess immunosuppressive activity, which may be useful for the treatment of neurological disorders of inflammatory etiology. Although the long-term safety of MSC-based therapies remains unclear, a large amount of both pre-clinical and clinical trials have shown functional improvements in animal models of nervous system diseases following transplantation of MSCs. In fact, there are several ongoing clinical trials evaluating the possible benefits this cell-based therapy could provide to patients with neurological damage, as well as their clinical limitations. In this review we focus on the potential of MSCs as a therapeutic tool to treat neurological disorders, summarizing the state of the art of this topic and the most recent clinical studies.