백서의 두개골 결손부에서 제대혈 유래 간엽줄기세포의 골재생에 관한 연구

이의석,장현석,권종진,이진용,임재석,서병무,윤정현,임재형 대한통합치과학회 2013 대한통합치과학회지 Vol.2 No.1

It has been suggested that umbilical cord blood (UCB) contains multipotent stem cells. Some authors suggest that within UCB are cells with potential for differentiation down mesenchymal lineages and UCB is one of the readily available sources of stem cells that may be used for bone regeneration. Isolation and characterization of these cells has been accomplished in some centers. Differentiation of these cells down multiple lineages has been documented. The use of UCB derived stem cells was investigated in this animal studies. Polycaprolactone‐20% tricalcium phosphate (PCL‐TCP) polymers allow for cellular adherence and differentiation and are adequate scaffolds for bone regeneration in tissue engineering strategies. The purpose of this study was to evaluate the potential of MSCs from UCB seeded onto PCL‐TCP composite scaffolds for bone tissue engineering. MSCs were isolated from UCB. MSCs from UCB seeded onto PCL‐TCP scaffolds were then implanted in calvarial bone defect of the athymic mouse. As a control, in each mouse was treated with scaffold only. At six and twelve weeks post‐implantation, histological examinations were performed. MSCs from UCB with PCL‐TCP scaffolds did not exhibited bone regeneration in calvarial bone defects in athymic mouse. Histological examination did not reveal the presence of regenerated bone of the defect. MSCs from UCB should be considered further applications in tissue‐engineered bone regeneration.

Mesenchymal Stem Cells Decrease Oxidative Stress in the Bowels of Interleukin-10 Knockout Mice

Kyong Jin Jung,Gun Woo Lee,Chul Hyun Park,Tae Jin Lee,Joo Young Kim,Eon Gi Sung,Seong Yong Kim,Byung Ik Jang,In Hwan Song 거트앤리버 소화기연관학회협의회 2020 Gut and Liver Vol.14 No.1

Background/Aims: Inflammatory bowel disease (IBD) is an autoimmune disease characterized by chronic inflammation mainly in the large intestine. The interleukin-10 knockout (IL-10 KO) mouse is a well-known animal model of IBD that develops spontaneous intestinal inflammation resembling Crohn’s disease. Oxidative stress is considered to be the leading cause of cell and tissue damage. Reactive oxygen species (ROS) can cause direct cell injury and/or indirect cell injury by inducing the secretion of cytokines from damaged cells. This study evaluated the effects of mesenchymal stem cell (MSC) on the progression of IBD. Methods: In this study, human bone marrow-derived MSCs were injected into IL-10 KO mice (MSC). Oxidative stress and inflammation levels were evaluated in the large intestine and compared with those in control IL-10 KO mice (CON) and normal wild-type control mice (Wild). Results: The levels of ROS (superoxide and hydrogen peroxidase) and a secondary end-product of lipid peroxidation (malondialdehyde) were considerably higher in the CON, while superoxide dismutase and catalase levels were lower in the MSC. Inflammation-related marker (interferon-γ, tumor necrosis factor-α, IL-4, and CD8) expression and inflammatory histological changes were much less pronounced in MSC than in CON. Conclusions: MSCs affect the redox balance, leading to the suppression of IBD.

개 지방세포 유래의 중간엽 줄기세포의 종양형성시험

이은선(Eun-Sun Lee),권은아(Euna Kwon),박정란(Jeong Ran Park),강병철(Byung-Chul Kang),강경선(Kyung-Sun Kang),조명행(Myung-Haing Cho) 한국독성학회 2007 Toxicological Research Vol.23 No.3

Several recent studies demonstrated the potential of bioengineering using stem cells in regenerative medicine. Adult mesenchymal stem cells (MSCs) have the pluripotency to differentiate into cells of mesodermal origin, i.e., bone, cartilage, adipose, and muscle cells; they, therefore, have many potential clinical applications. On the other hand, stem cells possess a self-renewal capability similar to cancer cells. For safety evaluation of MSCs, in this study, we tested tumorigenecity of canine adipose derived mesenchymal stem cells (cAD-MSCs) using Balb/c-nu mice. In this study, there were no changes in mortality, clinical signs, body weights and biochemical parameters of all animals treated. In addition, there were no significant changes between control and treated groups in autopsy findings. These results indicate that cAD-MSC has no tumorigenic potential under the condition in this study.

In Vitro Monitoring of Cardiomyogenic Differentiation of Mesenchymal Stem Cells using Sodium Iodide Symporter Gene

( Min Hwan Kim ),( Yong Jin Lee ),( Kwang Il Kim ),( Tae Sup Lee ),( Kwang Sun Woo ),( Dong Soo Lee ),( Chan Wha Kim ),( Chang Woon Choi ),( Sang Moo Lim ),( Joo Hyun Kang ) 한국조직공학·재생의학회 2012 조직공학과 재생의학 Vol.9 No.6

There is a need for non-invasive monitoring the differentiation of transplanted cell in stem cell therapy. In this study, we evaluated the usefulness of sodium iodide symporter (NIS) gene as molecular imaging reporter gene during cardiomyogenic differentiation of bone marrow derived mesenchymal stem cells (BMSCs). In a previous study, we showed that a transgenic mouse model expressing NIS driven by the α-myosin heavy chain (α-MHC) promoter was developed to image cardiomyocytes in vivo with a γ-camera and micro positron emission tomography (microPET). BMSCs were found to express stem cell specific surface markers including stem cell antigen-1 (Sca- 1) and CD44. After treatment with all-trans retinoic acid (ATRA), BMSCs morphologically changed and acquired a cardiomyocyte-like shape. The cardiomyogenic differentiation of BMSCs resulted in upregulated expression of cardiac specific genes including α-MHC and ventricular myosin light chain 2 (MLC-2v) and decreased expression of the stem cell specific marker, Sca-1. The ATRA treated BMSCs exhibited higher 125I uptake than that of nontreated cells, and this result suggested that the cardiomyogenic differentiation upregulated NIS gene activity as reporter in vitro. We demonstrated that, under the control of the α-MHC promoter, NIS activity reflected cardiomyogenic differentiation of BMSCs in vitro, and an imaging system based on NIS, the reporter gene, may be a useful tool for monitoring in the lineage specific differentiation of stem cells.

Optimization of Gene Delivery Mediated by Lipoplexes and Electroporation into Mouse Mesenchymal Stem Cells

Jong Chul Kim,Hong Sung Kim,Yeon Kyung Lee,Jung Seok Kim,Sang Il Park,Hwa Yeon Jung,Yong Serk Park 대한의생명과학회 2009 Biomedical Science Letters Vol.15 No.4

Recently, mesenchymal stem cells (MSCs) began to be utilized as a vehicle for ex vivo gene therapy based on their plasticity. Effective and safe transfection of therapeutic genes is a critical step for genetic modification of MSCs. Therefore, optimization of in vitro gene delivery into MSCs is essential to provide genetically modified stem cells. In this study, various cationic liposomes, O,O'-dimyristyl-N-lysyl aspartate (DMKD), DMKD/cholesterol, O,O'-dimyristyl- N-lysyl glutamate (DMKE), DMKE/cholesterol, and N-[1-(2,3-dioleoyloxy)]-N,N,N-trimethylammonium propane methyl sulfate (DOTAP)/cholesterol, were mixed with plasmid DNA encoding luciferase (pAAV-CMV-Luc) at varied ratios, and then used for transfection to MSCs under varied conditions. The MSCs were also transfected by electroporation under varied conditions, such as voltage, pulse length, and pulse interval. According to the experimental results, electroporation-mediated transfection was more efficient than cationic liposome-mediated transfection. The best MSC transfection was induced by electroporation 3 times pulses for 2 ms at 200 V with 10 seconds of a pulse interval.

Isolation of High Purity Mouse Mesenchymal Stem Cells through Depleting Macrophages Using Liposomal Clodronate

Song Ju Han,김정우,Lee Mi Nam,Oh Sin-Hye,Piao Xianyu,Wang Zhao,권승희,김옥수,고정태 한국조직공학과 재생의학회 2022 조직공학과 재생의학 Vol.19 No.3

BACKGROUND: The use of mouse bone marrow mesenchymal stem cells (mBMSCs) represents a promising strategy for performing preclinical studies in the field of cell-based regenerative medicine; however, mBMSCs obtained via conventional isolation methods have two drawbacks, i.e., (i) they are heterogeneous due to frequent macrophage contamination, and (ii) they require long-term culturing for expansion. METHODS: In the present study, we report a novel strategy to generate highly pure mBMSCs using liposomal clodronate. This approach is based on the properties of the two cell populations, i.e., BMSCs (to adhere to the plasticware in culture dishes) and macrophages (to phagocytose liposomes). RESULTS: Liposomal clodronate added during the first passage of whole bone marrow culture was selectively engulfed by macrophages in the heterogeneous cell population, resulting in their effective elimination without affecting the MSCs. This method allowed the generation of numerous high-purity Sca-1?CD44?F4/80- mBMSCs ([95%) with just one passaging. Comparative studies with mBMSCs obtained using conventional methods revealed that the mBMSCs obtained in the present study had remarkably improved experimental utilities, as demonstrated by in vitro multilineage differentiation and in vivo ectopic bone formation assays. CONCLUSION: Our newly developed method, which enables the isolation of mBMSCs using simple and convenient protocol, will aid preclinical studies based on the use of MSCs.

Hair growth promoting effect of dermal papilla like tissues from canine adipose-derived mesenchymal stem cells through vascular endothelial growth factor

LEE, Aeri,BAE, Sohee,LEE, Seung Hoon,KWEON, Oh-Kyeong,KIM, Wan Hee The Japanese Society of Veterinary Science 2016 The Journal of veterinary medical science Vol.78 No.12

<P>The purpose of this study was to investigate the protein expression pattern and the <I>in vivo</I> trichogenicity of dermal papilla-like tissues (DPLTs) made from canine adipose-derived mesenchymal stem cells (ASCs) in athymic nude mice. Canine ASCs were isolated and cultured from adipose tissue, and differentiation was induced by culturing ASCs in dermal papilla forming media. DPLTs were embedded in collagen gel, and their structural characteristics and protein expression were evaluated by hematoxylin and eosin stain and immunohistochemistry. Athymic nude mice were divided into two groups (control and DPLTs groups), and DPLTs were injected in skin wounds of mice in the DPLTs group. The trichogenicity of DPLTs was assessed by gross and histological evaluations for 30 days. The fate and the growth factor-secretion effect of DPLTs were evaluated by immunohistochemistry and Western blotting. DPLTs have a compact aggregated structure, form extracellular matrix and highly express the protein specific for dermal papillae, including ALP and versican. New hair follicle formation was remarkable in nude mice of the DPLTs group in gross findings and H&E stain. Vascularization was increased in the DPLTs group, which was the effect of vascular endothelial growth factor secreted by DPLTs <I>in vitro</I> and <I>in vivo</I>. These data suggest that engineered canine DPLTs have characteristics of dermal papillae and have a positive effect on hair regeneration by secreting growth factors.</P>

The effect of embryogenic stem cell-derived mesenchymal stem cells on imiquimod-induced psoriasis-like skin inflammation in mice

( Na Gyeong Yang ),( Tae Heum Park ),( Dong Sung Kim ),( Eun Ji Hong ),( Hee Jung Yoon ),( Sung Yul Lee ),( Yu Sung Choi ),( Young Lip Park ),( Man Ryul Lee ),( Jung Eun Kim ) 대한피부과학회 2022 대한피부과학회 학술발표대회집 Vol.74 No.1

Effects of Human Adipose-derived Stem Cells on Cutaneous Wound Healing in Nude Mice

( Seung Ho Lee ),( Joon Ho Lee ),( Kwang Hyun Cho ) 대한피부과학회 2011 Annals of Dermatology Vol.23 No.2

Background: Despite numerous treatments available for deteriorated cutaneous wound healing such as a diabetic foot, there is still the need for more effective therapy. Adipose-derived stem cells (ASCs) are mesenchymal stem cells, which are self-renewing and multipotent. Mesenchymal stem cells have the potential for tissue repair and regeneration. Objective: To investigate the effects of human ASCs on the healing of cutaneous wounds in nude mice. Methods: 15-mm round full-thickness skin defects were generated on the back of BALB/c nude mice. The mice were divided into three groups for wound coverage: (i) human ASCs-populated collagen gel, (ii) human dermal fibroblasts- populated collagen gel, and (iii) collagen gel alone. Wound contraction was prevented with a splint method. Wound size was measured 10 days after injury. At 28 days histological analysis was performed. Results: Both ASCs and dermal fibroblasts accelerated wound closure, but dermal fibroblasts were more effective than ASCs. At 28 days, the dermal portion of ASCs or dermal fibroblasts wound scars were thicker than collagen gel wound scars. Conclusion: ASCs and dermal fibroblasts stimulate cutaneous wound healing and improve scar thickness. (Ann Dermatol 23(2) 150∼155, 2011)

The effect of human mesenchymal stem cell injection on pain behavior in chronic post-ischemia pain mice

유시현,Sung Hyun Lee,Seunghwan Lee,Jae Hong Park,Seunghyeon Lee,Heecheol Jin,Hue Jung Park 대한통증학회 2020 The Korean Journal of Pain Vol.33 No.1

Background: Neuropathic pain (NP) is considered a clinically incurable condition despite various treatment options due to its diverse causes and complicated disease mechanisms. Since the early 2000s, multipotent human mesenchymal stem cells (hMSCs) have been used in the treatment of NP in animal models. However, the effects of hMSC injections have not been studied in chronic post-ischemia pain (CPIP) mice models. Here, we investigated whether intrathecal (IT) and intrapaw (IP) injections of hMSCs can reduce mechanical allodynia in CPIP model mice. Methods: Seventeen CPIP C57/BL6 mice were selected and randomized into four groups: IT sham (n = 4), IT stem (n = 5), IP sham (n = 4), and IP stem (n = 4). Mice in the IT sham and IT stem groups received an injection of 5 μL saline and 2 × 104 hMSCs, respectively, while mice in the IP sham and IP stem groups received an injection of 5 μL saline and 2 × 105 hMSCs, respectively. Mechanical allodynia was assessed using von Frey filaments from pre-injection to 30 days post-injection. Glial fibrillary acidic protein (GFAP) expression in the spinal cord and dorsal root ganglia were also evaluated. Results: IT and IP injections of hMSCs improved mechanical allodynia. GFAP expression was decreased on day 25 post-injection compared with the sham group. Injections of hMSCs improved allodynia and GFAP expression was decreased compared with the sham group. Conclusions: These results suggested that hMSCs may be also another treatment modality in NP model by ischemia-reperfusion.