• Archives of Plastic Surgery
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• 제37권4호
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• pp.323-328
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• 2010

Purpose: Adipose-derived stromal cells (ADSCs) are multipotent cells that have been found to promote wound healing through the process of angiogenesis and reepithelialization. Generally, it is well known that the antigenicity of ADSCs doesn't affect stem cell therapy. In this study, we investigated the effect of allogeneic ADSCs in the wound healing process by applying allogeneic ADSCs on the wound healing splint model of mice. Methods: Adipose tissue was harvested from the epididymal fat pads of BALB/c and C57BL/6 mice. Twenty four mice BALB/c were divided into three groups; control, isogeneic, and allogeneic groups. Two full thickness defects with 6 mm diameters were created on the back of BALB/c mice. $1{\times}10^6$ ADSCs from BALB/c mice were applied on the isogeneic group. In the allogeneic group, ADSCs from the C57BL/6 mice were applied. No cells were applied to the control group. The sizes of the wounds were evaluated in 3, 5, 7, 10, and 14 days after the wounds were applied, and tissues were harvested in 7 and 14 days for histological analysis. Results: Wound healing rates had showed significant increase in 10, and 14 days when the isogeneic group was compared to the control group, but the allogeneic group showed significantly decrease compared to the isogeneic group (p<0.05). Histological scores in the isogeneic group were significantly high, but significantly lower in the allogeneic group when compared to the isogeneic group in 2 weeks (p<0.05). In the isogeneic group, thick inflammatory cell infiltration with abundant capillaries were observed in 1 week, and thick epithelium with many large capillaries were observed in 2 weeks. Conclusion: When isogeneic ADSCs were applied to wounds, they presented a faster wound healing rate compared to controls and the allogeneic group. Unlike general stem cell therapy, these findings suggest that cell therapy targeted at enhancing wound healing may benefit from the use of ADSCs with identical antigenicity, as opposed to allogeneic or xenogenic ADSCs.

• 생명과학회지
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• 제31권9호
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• pp.856-861
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• 2021

CD82/KAI1은 분자촉진제로서 암 전이억제자로 역할이 잘 알려져 있으나, 최근 줄기 전구 세포와 혈관 신생, 근육에서 다양한 역할들이 밝혀지고 있다. 이에 본 연구진은 최근에 보고된 CD82의 다양한 기능과 역할에 관하여 총설 하고자 한다. CD82는 4개의 막 통과 도메인을 가진 테트라스파닌의 한 종류로 암의 전이 과정에 관여하는 세포접착분자들과의 상호작용을 통하여 암세포의 이동 능력을 저해한다. 암 전이 억제자로의 기능 외에도 줄기세포 니쉬에서도 그 역할이 밝혀졌다. 골수에서 분화재생능력이 뛰어난 최상위 조혈모세포(LT-HSC)에서 CD82가 발현되며, DARC와의 상호결합으로 줄기세포의 휴면을 유도한다. 줄기세포의 휴면 조절 외에도, CD82는 Rac1 활성 조절을 통해 조혈모세포의 골수로의 귀환 및 생착에도 역할을 한다. 또한, CD82는 근육 줄기 세포의 분화능을 유지시키며, 혈관 내피세포에서 세포 접착 분자와 IL-6, VEGF와 같은 사이토카인의 발현을 저해하여 혈관 신생을 억제한다. CD82는 다양한 조직 및 줄기-전구 세포에서 계급을 구별할 수 있는 핵심 세포막 표면 단백질이며, 세포 자원의 증폭 및 검증에 있어 중요하다. 다양한 기관과 세포에서 CD82의 역할과 추가적인 연구들이 줄기세포치료의 임상적 적용에 있어 큰 도움이 되기를 기대한다.

• BMB Reports
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• 제52권5호
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• pp.318-323
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• 2019

Mesenchymal stem cells (MSCs) are multipotent adult stem cells that present immunosuppressive effects in experimental and clinical trials targeting various rare diseases including inflammatory bowel disease (IBD). In addition, recent studies have reported tryptophanyl-tRNA synthetase (WRS) possesses uncanonical roles such as angiostatic and anti-inflammatory effects. However, little is known about the function of WRS in MSC-based therapy. In this study, we investigated if a novel factor, WRS, secreted from MSCs has a role in amelioration of IBD symptoms and determined a specific mechanism underlying MSC therapy. Experimental colitis was induced by administration of 3% DSS solution to 8-week-old mice and human umbilical cord blood-derived MSCs (hUCB-MSCs) were injected intraperitoneally. Secretion of WRS from hUCB-MSCs and direct effect of WRS on isolated $CD4^+$ T cells was determined via in vitro experiments and hUCB-MSCs showed significant therapeutic rescue against experimental colitis. Importantly, WRS level in serum of colitis induced mice decreased and recovered by administration of MSCs. Through in vitro examination, WRS expression of hUCB-MSCs increased when cells were treated with interferon-${\gamma}$ ($IFN-{\gamma}$). WRS was evaluated and revealed to have a role in inhibiting activated T cells by inducing apoptosis. In summary, $IFN-{\gamma}$-mediated secretion of WRS from MSCs has a role in suppressive effect on excessive inflammation and disease progression of IBD and brings new highlights in the immunomodulatory potency of hUCB-MSCs.

• 생명과학회지
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• 제21권5호
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• pp.631-646
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• 2011

중간엽 줄기 세포는 다분화능을 가지고 있으며 골수, 지방, 태반, 치아속질, 윤활막, 편도 및 가슴샘 등 인체의 다양한 조직에서 분리된다. 중간엽 줄기세포는 조직의 항상성을 조절하며 다분화능, 분리와 조작의 용이함, 암세포로의 화학주성 및 면역 반응 조절 등의 특징을 가지고 있어서 재생 의학, 암 치료 및 식대주 질환(GVHD) 등에 이용할 수 있는 세포치료제로 주목 받고 있다. 하지만 주위 세포와 조직을 지지하고 조절하는 특징과 관련하여 중간엽 줄기세포가 혈관 생성을 촉진하고 성장인자를 분비하며 암세포를 공격하는 면역 반응을 억제함으로써 암의 진행을 촉진시킨다는 사실 또한 보고 되고 있다. 이러한 사실들로 인해 중간엽 줄기세포의 임상 적용이 제한되고 있다. 본 연구에서는 어떠한 기전을 통해서 중간엽 줄기세포가 암의 진행을 촉진하는 지지 세포로 기능하는지를 밝히기 위해서 인체 지방 조직에서 유래한 중간엽 줄기세포를 두 개의 암세포주(H460, U87MG)와 각각 공동 배양하고 microarray를 이용해서 암세포와 공동 배양되지 않은 중간엽 줄기세포와 유전자의 발현을 비교하였다. 두 암세포주와 공동배양에서 공통적으로 2배 이상 차이 나는 유전자를 DAVID (Database for Annotation, Visualization and Integrated Discovery)와 PANTHER (Protein ANalysis THrough Evolutionary Relationships)를 이용해 분석하였으며 생물학적 과정, 분자적 기능, 세포의 구성 성분, 단백질의 종류, 질병과 인체 조직 그리고 신호전달에 관련된 정보를 획득하였다. 이를 통해서 암세포는 중간엽 줄기세의 분화, 증식, 에너지 대사, 세포의 구조 및 분비기능을 조절하여 유전자의 발현 양상을 암 연관 섬유모세포(cancer associated fibroblast)와 유사한 세포로 변형 시킨다는 사실을 알 수 있었다. 본 연구의 결과는 중간엽 줄기세포를 이용한 임상 치료제의 효과와 안정성을 개선하는데 응용될 수 있을 것이다.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제32권4호
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• pp.299-305
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• 2010

Purpose: Adipose tissue is located beneath the skin, around internal organs, and in the bone marrow in humans. Its main role is to store energy in the form of fat, although it also cushions and insulates the body. Adipose tissue also has the ability to dynamically expand and shrink throughout the life of an adult. Recently, it has been shown that adipose tissue contains a population of adult multipotent mesenchymal stem cells and endothelial progenitor cells that, in cell culture conditions, have extensive proliferative capacity and are able to differentiate into several lineages, including, osteogenic, chondrogenic, endothelial cells, and myogenic lineages. Materials and Methods: This study focused on endothelial cell culture from the adipose tissue. Adipose tissues were harvested from buccal fat pad during bilateral sagittal split ramus osteotomy for surgical correction of mandibular prognathism. The tissues were treated with 0.075% type I collagenase. The samples were neutralized with DMEM/and centrifuged for 10 min at 2,400 rpm. The pellet was treated with 3 volume of RBC lysis buffer and filtered through a 100 ${\mu}m$ nylon cell strainer. The filtered cells were centrifuged for 10 min at 2,400 rpm. The cells were further cultured in the endothelial cell culture medium (EGM-2, Cambrex, Walkersville, Md., USA) supplemented with 10% fetal bovine serum, human EGF, human VEGF, human insulin-like growth factor-1, human FGF-$\beta$, heparin, ascorbic acid and hydrocortisone at a density of $1{\times}10^5$ cells/well in a 24-well plate. Low positivity of endothelial cell markers, such as CD31 and CD146, was observed during early passage of cells. Results: Increase of CD146 positivity was observed in passage 5 to 7 adipose tissue-derived cells. However, CD44, representative mesenchymal stem cell marker, was also strongly expressed. CD146 sorted adipose tissue-derived cells was cultured using immuno-magnetic beads. Magnetic labeling with 100 ${\mu}l$ microbeads per 108 cells was performed for 30 minutes at $4^{\circ}C$ a using CD146 direct cell isolation kit. Magnetic separation was carried out and a separator under a biological hood. Aliquous of CD146+ sorted cells were evaluated for purity by flow cytometry. Sorted cells were 96.04% positivity for CD146. And then tube formation was examined. These CD146 sorted adipose tissue-derived cells formed tube-like structures on Matrigel. Conclusion: These results suggest that adipose tissue-derived cells are endothelial cells. With the fabrication of the vascularized scaffold construct, novel approaches could be developed to enhance the engineered scaffold by the addition of adipose tissue-derived endothelial cells and periosteal-derived osteoblastic cells to promote bone growth.

• 한국발생생물학회지:발생과생식
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• 제11권3호
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• pp.235-243
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• 2007

인간 제대혈 세포는 조혈모세포, 중간엽 줄기세포와내피전구세포를 풍부하게 포함하고 있다. 인간 제대혈 속의 중간엽 줄기세포는 조혈모세포와는 달리 다능성 줄기세포이며 신경세포로 분화할 수 있는 잠재성을 가지고 있다. 본 연구에서는 세포배양을 통해 제대혈의 중간엽 줄기세포를 신경세포와 콜린성 신경세포로 분화를 유도하였다. 중간엽 줄기세포를 신경세포로 분화시키기 위해 배양액에 dimethyl sulphoxide(DMSO)와 butylated hydroxyanisole(BHA)를 첨가하여 유도하였으며 basic fibroblast growth factor(bFGF), retinoic acid(RA), sonic hedgehog(Shh)를 처리하여 콜린성 신경세포로 분화시켰다. DMSO와 BHA에 처리된 중간엽 줄기세포가 빠르게 신경세포 모양으로 분화하는 것을 관찰하였으며, 이것은 면역조직학적 염색에서 신경세포 특이 표지인 $\beta$-tubulin III, 별아교세포에 대한 특이 표지인 GFAP, 희돌기아교세포에 대한 특이 표지인 Gal-C에 대해 양성반응을 나타내었고, 그 비율은 각각 $32.3{\pm}2.9%$, $11.0{\pm}0.9%,\;9.4{\pm}1.0%$였다. RT-PCR 분석에서 배양 단계에 따라 신경세포에 특이적인 표지 인자가 발현됨을 통해, 중간엽 줄기세포가 신경세포로 분화됨을 확인하였다. 또한, 중간엽 줄기세포에 bFGF, RA, Shh를 처리하여 콜린성 신경세포로 분화시켰을 때, 전체 중간엽 세포 중 $31.3{\pm}3.2%$가 신경세포 특이 표지인 $\beta$-tubulin III에 양성반응을 보였으며 이들 세포 중 $70.0{\pm}7.8%$가 콜린성 신경 특이 표지인 ChAT에 양성반응을 보였고, 이것은 Woodbury 방법에 의한 신경분화의 경우보다 3배 가량 높은 비율로 콜린성 신경의 분화를 유도한 것이다. 이러한 실험 결과들은 인간 제대혈의 중간엽 줄기세포가 콜린성 신경세포로 분화가 가능하고 이러한 잠재성을 가진 제대혈 중간엽 줄기세포는 퇴행성 신경질환에 대한 세포 치료제로서 가능성을 제시한다.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제36권3호
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• pp.186-196
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• 2010

Introduction: The first aim of this study was to isolate the dental tissue-derived stem cells from the dental follicle (DF), dental pulp (DP), and root apical papilla (RAP) of the extracted wisdom teeth. Second was to evaluate their characterization with the expressions of transcription factors and cell surface markers. Finally, their ability of the in vitro multi-lineage differentiations into osteogenic and adipogenic cells were compared, respectively. Materials and Methods: Dental tissues, including dental follicle, dental pulp, and root apical papilla, were separated in the extracted wisdom teeth. These three dental tissues were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with supplements, respectively. After passage 3, the homogeneous shaped dental tissue-derived cells were analyzed the expression of transcription factors (Oct-4, Nanog and Sox-2) and cell surface markers (CD44, CD90 and CD105) with reverse transcription polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. In order to evaluate in vitro multi-lineage differentiations, the culture media were changed to the osteogenic and adipogenic induction mediums when the dental tissue-derived cells reached to passage 3. The characteristics of these three dental tissue-derived cells were compared with immunohistochemistry. Results: During primary culture, heterogenous and colony formatted dental tissue-derived cells were observed in the culture plates. After passage 2 or 3, homogenous spindle-like cells were observed in all culture plates. Transcription factors and mesenchymal stem cell markers were positively observed in all three types of dental tissue-derived cells. However, the quantity of expressed transcription factors was most large in RAP-derived cells. In all three types of dental tissue-derived cells, osteogenic and adipogenic differentiations were observed after treatment of specific induction media. In vitro adipogenic differentiation was similar among these three types of cells. In vitro osteogenic differentiation was most strongly and frequently observed in the RAP-derived cells, whereas rarely osteogenic differentiation was observed in the DP-derived cells. Conclusion: These findings suggest that three types of human dental tissue-derived cells from extracted wisdom teeth were multipotent mesenchymal stem cells, have the properties of multi-lineage differentiations. Especially, stem cells from root apical papilla (SCAP) have much advantage in osteogenic differentiation, whereas dental follicle cells (DFCs) have a characteristic of easy adipogenic differentiation.