Human Tissue-Engineered Skeletal Muscle: A Tool for Metabolic Research

김지훈,유승민,손장원 대한내분비학회 2022 Endocrinology and metabolism Vol.37 No.3

Skeletal muscle is now regarded as an endocrine organ based on its secretion of myokines and exerkines, which, in response to metabolic stimuli, regulate the crosstalk between the skeletal muscle and other metabolic organs in terms of systemic energy homeostasis. This conceptual basis of skeletal muscle as a metabolically active organ has provided insights into the potential role of physical inactivity and conditions altering muscle quality and quantity in the development of multiple metabolic disorders, including insulin resistance, obesity, and diabetes. Therefore, it is important to understand human muscle physiology more deeply in relation to the pathophysiology of metabolic diseases. Since monolayer cell lines or animal models used in conventional research differ from the pathophysiological features of the human body, there is increasing need for more physiologically relevant in vitro models of human skeletal muscle. Here, we introduce recent studies on in vitro models of human skeletal muscle generated from adult myogenic progenitors or pluripotent stem cells and summarize recent progress in the development of three-dimensional (3D) bioartificial muscle,which mimics the physiological complexity of native skeletal muscle tissue in terms of maturation and functionality. We then discussthe future of skeletal muscle 3D-organoid culture technology in the field of metabolic research for studying pathological mechanisms and developing personalized therapeutic strategies.

In Vitro and In Vivo Osteogenesis of Human Orbicularis Oculi Muscle-Derived Stem Cells

Guangpeng Liu,Caihe Liao,Xi Chen,Yaohao Wu 한국조직공학과 재생의학회 2018 조직공학과 재생의학 Vol.15 No.4

BACKGROUND: Cell-based therapies for treating bone defects require a source of stem cells with osteogenic potential. There is evidence from pathologic ossification within muscles that human skeletal muscles contain osteogenic progenitor cells. However, muscle samples are usually acquired through a traumatic biopsy procedure which causes pain and morbidity to the donor. Herein, we identified a new alternative source of skeletal muscle stem cells (SMSCs) without conferring morbidity to donors. METHODS: Adherent cells isolated from human orbicularis oculi muscle (OOM) fragments, which are currently discarded during ophthalmic cosmetic surgeries, were obtained using a two-step plating method. The cell growth kinetics, immunophenotype and capabilities of in vitro multilineage differentiation were evaluated respectively. Moreover, the osteogenically-induced cells were transduced with GFP gene, loaded onto the porous b-tricalcium phosphate (b-TCP) bioceramics, and transplanted into the subcutaneous site of athymic mice. Ectopic bone formation was assessed and the cell fate in vivo was detected. RESULTS: OOM-derived cells were fibroblastic in shape, clonogenic in growth, and displayed phenotypic and behavioral characteristics similar to SMSCs. In particular, these cells could be induced into osteoblasts in vitro evidenced by the extracellular matrix calcification and enhanced alkaline phosphatase (ALP) activity and osteocalcin (OCN) production. New bone formation was found in the cell-loaded bioceramics 6 weeks after implantation. By using the GFP-labeling technique, these muscle cells were detected to participate in the process of ectopic osteogenesis in vivo. CONCLUSION: Our data suggest that human OOM tissue is a valuable and noninvasive resource for osteoprogenitor cells to be used in bone repair and regeneration.

Rejuvenation of Aged Skeletal Muscle by Systematic Factor, GDF11

( Young C. Jang ),( Manisha Sinha ),( Amy J. Wagers ) 한국체육학회 2015 국제스포츠과학 학술대회 Vol.2015 No.1

Purpose: Recent studies involving heterochronic parabiosis indicate that impaired skeletal muscle regeneration in aged animals is in part systemically controlled and reversible by exposure to a young circulation. While prior studies have identified a handful of systemic "aging" factors, discovery of the humoral "rejuvenating" factors that act on muscle stem cells (satellite cells) to restore regenerative function has been relatively more elusive. Here, we demonstrate that the circulating hormone Growth Differentiation Factor 11 (GDF11) is a rejuvenating factor for skeletal muscle. Methods: Using either heterochronic parabiosis (young and aged mice with shared circulation) or systemic delivery of recombinant GDF11 (0.1 mg/KG), in vitro and in vivo muscle stem cell function, regenerative potential, as well as overall muscle function were assessed. Results: Supplementation of systemic GDF11 levels, which normally decline with age, is sufficient to reverse functional impairments and restore genomic integrity in skeletal muscle stem cells. Augmentation of GDF11 levels further improved structural and functional features of resting skeletal muscle, resulting in increased strength and enhanced endurance exercise capacity. Conclusion: Taken together, these data reveal critical mechanisms in the systemic regulation of aging and identify a promising candidate therapeutic for the reversal of age-related skeletal muscle and stem cell dysfunction.

흰쥐 부분 근육 절제 모델에서 지방유래줄기세포의 치료 효과

황옥경 ( Ok Kyung Hwang ),홍일화 ( Il Hwa Hong ),기미란 ( Mi Ran Ki ),( Se Il Park ),박진규 ( Jin Kyu Park ),한정연 ( Jung Youn Han ),홍경숙 ( Kyung Sook Hong ),지애리 ( Ae Ri Ji ),정재호 ( Jae Ho Jeong ),강경선 ( Kyung Seon Ka 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.4

Adipose tissue derived stem cells(ASCs) are multipotent cells capable of forming bone, cartilage, and other connective tissues. In this study we evaluated the effect of stem cells derived from adipose tissue on the muscle regeneration in partial removed skeletal muscle in rats. Male sprague-dawley rats were randomly divided into three groups: control, PBS-injected and ASCs-injected group. Stem cells were isolated from rat abdominal adipose tissue by collagenase digestion and ASCs labeled with superparamagnetic iron oxide(Feridex) were used. Muscle injury was subjected to rat bypartial removal of quadriceps femoris muscle. The labeled cells were directly injected into the muscle injured area after muscle injury an hour. After 2 weeks, histological analysis was performed using H&E staining and prussian blue staining. The muscle regeneration markers were examined by immunohistochemistry and RT-PCR. ASCs labeled with Feridex were detected by prussian blue staining in injured muscle area. Microscopical muscle regeneration and neovascularization were more increased in ASCs-injected group compared to PBS-injected group. The expression level of Pax7, MyoD and myogenin were more significantly increased in ASCs-injected group compared with the PBS-injected group. This preliminary observation shows that ASCs may be directly involved in the regeneration process of injured skeletal muscle; therefore the possibility of using ASCs as a stem cell therapy for muscle regeneration need to be further investigated for the purpose of clinical trials.

Unraveling the Paradoxical Action of Androgens on Muscle Stem Cells

서지윤,김지훈,공영윤 한국분자세포생물학회 2019 Molecules and cells Vol.42 No.2

Androgens act in almost all tissues throughout the lifetime and have important roles in skeletal muscles. The levels of androgens increase during puberty and remain sustained at high levels in adulthood. Because androgens have an anabolic effect on skeletal muscles and muscle stem cells, these increased levels of androgens after puberty should lead to spontaneous muscle hypertrophy and hyperplasia in adulthood. However, the maintenance of muscle volume, myonuclei number per myofiber, and quiescent state of satellite cells in adulthood despite the high levels of androgens produces paradoxical outcomes. Our recent study revealed that the physiological increase of androgens at puberty initiates the transition of muscle stem cells from proliferation to quiescence by the androgen-Mindbomb1-Notch signaling axis. This newly discovered androgen action on skeletal muscles underscores the physiological importance of androgens on muscle homeostasis throughout life. This review will provide an overview of the new androgen action on skeletal muscles and discuss the paradoxical effects of androgens suggested in previous studies.

Unraveling the Paradoxical Action of Androgens on Muscle Stem Cells

Seo, Ji-Yun,Kim, Ji-Hoon,Kong, Young-Yun Korean Society for Molecular and Cellular Biology 2019 Molecules and cells Vol.42 No.2

Androgens act in almost all tissues throughout the lifetime and have important roles in skeletal muscles. The levels of androgens increase during puberty and remain sustained at high levels in adulthood. Because androgens have an anabolic effect on skeletal muscles and muscle stem cells, these increased levels of androgens after puberty should lead to spontaneous muscle hypertrophy and hyperplasia in adulthood. However, the maintenance of muscle volume, myonuclei number per myofiber, and quiescent state of satellite cells in adulthood despite the high levels of androgens produces paradoxical outcomes. Our recent study revealed that the physiological increase of androgens at puberty initiates the transition of muscle stem cells from proliferation to quiescence by the androgen-Mindbomb1-Notch signaling axis. This newly discovered androgen action on skeletal muscles underscores the physiological importance of androgens on muscle homeostasis throughout life. This review will provide an overview of the new androgen action on skeletal muscles and discuss the paradoxical effects of androgens suggested in previous studies.

사람 탯줄혈액으로부터 다분화능 중간엽 줄기세포의 분리 및 분화

김호연,양일호,홍승현,장은지,정주아,황수한,김성환,안치영,한훈 한국생체재료학회 2004 생체재료학회지 Vol.8 No.1

Human umbilical cord blood (UCB) has been regarded as an alternative source for cell transplantation and cell therapy due to its hematopoietic potential of generating all blood cells, and mesenchymal potential of differentiating into various cells of a mesodermal origin. In this study, we pursued to isolate mesenchymal stem cells (MSCs) from UCB and to induce them to differentiate into possibly many tissue-specific cell types. During the cell culture expansion, UCB-derived mononuclear cells gave rise to adherent layers of fibroblast-like cells expressing MSCs-related antigens such as SH2, SH3, ASMA, CD13, CD29, and CD49e. Moreover, the study showed that these cells, in appropriate differentiation conditions, could differentiate, at least, into skeletal myoblasts, osteoblasts, and neuronal cells, as judged by RT-PCR, immunocytochemistry, and immunofluorescent staining against several tissue-specific markers. Our results indicate that UCB may be an alternative source of MSCs to bone marrow as UCB-derived MSCs possess a multipotency of skeletal myogenic, osteogenic, and neurogenic differentiation. And it also implies that these cells may become a suitable source for cell-based therapy as well as a useful tool of tissue engineering.

Myogenic differentiation potential of human tonsil-derived mesenchymal stem cells and their potential for use to promote skeletal muscle regeneration

PARK, SAEYOUNG,CHOI, YOONYOUNG,JUNG, NAMHEE,YU, YEONSIL,RYU, KYUNG-HA,KIM, HAN SU,JO, INHO,CHOI, BYUNG-OK,JUNG, SUNG-CHUL UNKNOWN 2016 INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE Vol.37 No.5

<P>Stem cells are regarded as an important source of cells which may be used to promote the regeneration of skeletal muscle (SKM) which has been damaged due to defects in the organization of muscle tissue caused by congenital diseases, trauma or tumor removal. In particular, mesenchymal stem cells (MSCs), which require less invasive harvesting techniques, represent a valuable source of cells for stem cell therapy. In the present study, we demonstrated that human tonsil-derived MSCs (T-MSCs) may differentiate into myogenic cells <I>in vitro</I> and that the transplantation of myoblasts and myocytes generated from human T-MSCs mediates the recovery of muscle function <I>in vivo</I>. In order to induce myogenic differentiation, the T-MSC-derived spheres were cultured in Dulbecco's modified Eagle's medium/nutrient mixture F-12 (DMEM/F-12) supplemented with 1 ng/ml transforming growth factor-β, non-essential amino acids and insulin-transferrin-selenium for 4 days followed by culture in myogenic induction medium [low-glucose DMEM containing 2% fetal bovine serum (FBS) and 10 ng/ml insulin-like growth factor 1 (IGF1)] for 14 days. The T-MSCs sequentially differentiated into myoblasts and skeletal myocytes, as evidenced by the increased expression of skeletal myogenesis-related markers [including α-actinin, troponin I type 1 (TNNI1) and myogenin] and the formation of myotubes <I>in vitro</I>. The <I>in situ</I> transplantation of T-MSCs into mice with a partial myectomy of the right gastrocnemius muscle enhanced muscle function, as demonstrated by gait assessment (footprint analysis), and restored the shape of SKM without forming teratomas. Thus, T-MSCs may differentiate into myogenic cells and effectively regenerate SKM following injury. These results demonstrate the therapeutic potential of T-MSCs to promote SKM regeneration following injury.</P>

Muscle stem cell senescence, muscle regeneration and Senolytics

Dong Ryeol Ryu(류동렬) 한국실험동물학회 2021 한국실험동물학회 학술발표대회 논문집 Vol.2021 No.1

동물생명공학 : DNA Microarray 분석을 통한 한우 부위별 특이 마커 유전자의 발굴

이은주 ( Eun Ju Lee ),이용석 ( Yong Seok Lee ),이현정 ( Hyun Jeong Lee ),최인호 ( In Ho Choi ),윤두학 ( Du Hak Yoon ),신유미 ( Yu Mi Shin ),전태훈 ( Tae Hoon Chun ) 한국동물자원과학회 ( 구 한국축산학회 ) 2010 한국축산학회지 Vol.52 No.4