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손미영 ( Mi Yeung Sohn ),서지민 ( Ji Min Seo ),조현정 ( Hyun Jung Cho ),서장수 ( Jang Soo Suh ),성원준 ( Won Joon Seong ),박일수 ( Il Soo Park ),( Anthony Atala ),유지 ( James J Yoo ),손윤희 ( Yun Hee Shon ) 한국조직공학·재생의학회 2012 조직공학과 재생의학 Vol.9 No.1s
Human stem cells are typically obtained for clinical application of cell therapy by isolating and subculturing adherent cells. The expansion of stem cells depends on the culture conditions and requires medium supplemented with fetal bovine serum (FBS) to generate relevant numbers of cells. However, the presence of FBS is a major obstacle for clinical use of the stem cells. Therefore, we have evaluated the capacity of expansion of amniotic fluid-derived stem cells (AFSCs) in a commercial serum-free medium in comparison with a conventional FBS-containing medium. AFSCs were expanded in two different culture media. Proliferation rate of the cells was higher in serum-free medium than in FBS-containing medium. Stem cells isolated in the both media were morphologically similar, and they were expressed identical surface antigens and mRNA of stem cell markers. AFSCs in the both media were also able to differentiate into the myogenic cells. These results suggest that a commercial serum-free medium (MesemCult-XF Medium) is an optimal medium for expansion of AFSCs. It allows a better cell expansion, preserves cell characteristics, and reduces the culture period. This medium seems suitable for clinical scale expansion of AFSCs.
( Hyojin Kim ),( Sun Young Moon ),( Mi Yeung Sohn ),( Weon Ju Lee ) 대한피부과학회 2017 Annals of Dermatology Vol.29 No.1
Background: Acne vulgaris has been linked to the Western diet. Hyperglycemic diet increases insulin and insulin-like growth factor (IGF)-1. Deeper insights into IGF-1-mediated signal pathway are critical importance to understand the impact of Western diet. Objective: We investigated the effect of IGF-1 on the expression of inflammatory biomarkers and sebum production in cultured sebocytes. Methods: Polymerase chain reaction and enzyme-linked immunosorbent assay were performed to measure changes in the expression of inflammatory biomarkers including interleukin (IL)-1β, IL-6, IL-8, tumor necrosis factor (TNF)-α, nuclear factor kappa- light-chain-enhancer of activated B cells (NF-κB), IGF1R, IGFBP2, sterol response element-binding protein (SREBP), and phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PI3KCA) after the treatment of cultured sebocytes with 10<sup>-7 </sup>M or 10<sup>-5</sup> M IGF-1. Sebum production was evaluated after the treatment of cultured sebocytes with 10<sup>-7</sup> M or 10<sup>-5</sup> M IGF-1 using lipid analysis. Results: The expression levels of IL-1β, IL-6, IL-8, and TNF-α in cultured sebocytes after treatment with 10<sup>-7</sup> M or 10<sup>-5</sup> M IGF-1 were increased. Increased gene expression levels of NF-κB in cultured sebocytes were also shown after 10<sup>-7</sup> M or 10<sup>-5</sup> M IGF-1 treatments. Gene expression of these inflammatory biomarkers was decreased after 10<sup>-7</sup> M or 10<sup>-5</sup> M IGF-1 treatment in the presence of 100 nM NF-κB inhibitor. Treatment with 10<sup>-7</sup> M or 10<sup>-5</sup> M IGF-1 increased the gene expression levels of IGF1R, IGFBP2, SREBP and PI3KCA in cultured sebocytes. Sebum production from cultured sebocytes treated with 10<sup>-7</sup> Mor 10<sup>-5</sup> M IGF-1 was also increased. Conclusion: It is suggestive that IGF-1 might be involved in the pathogenesis of acne by increasing both expression of inflammatory biomarkers and also sebum production in sebocytes. (Ann Dermatol 29(1) 20∼25, 2017)
Vitamin D increases expression of cathelicidin in cultured sebocytes.
Lee, Weon Ju,Cha, Hyun Wuk,Sohn, Mi Yeung,Lee, Seok-Jong,Kim, Do Won Springer-Verlag 2012 Archives of dermatological research Vol.304 No.8
<P>Antimicrobial peptides (AMPs) have been widely accepted as major contributors to cutaneous innate non-specific immunity, which is the first line of defense against invading pathogens. Altered expression of AMPs has been described in chronic inflammatory disorders of the skin. This study was conducted to clarify the expression of AMPs, including psoriasin, human β-defensin (hBD)-2, hBD-3, and LL-37, in cultured sebocytes and to evaluate the changes in the expression of AMPs in cultured sebocytes after treatment with vitamin D. Primary culture of sebocytes obtained from hair follicles of the scalp was used for this study. Cultured sebocytes were treated with vitamin D (10(-10)-10(-6)?M). In addition, vitamin D (10(-10), 10(-8) and 10(-6)?M) was added to sebocytes treated with vitamin D receptor (VDR) siRNA. Reverse transcription-polymerase chain reaction for psoriasin, hBD-2, hBD-3, and hCAP-18 was performed using cultured sebocytes and sebocytes treated with VDR siRNA. Western blotting for psoriasin, hBD-2, hBD-3, and hCAP-18 was also performed using sebocytes treated with vitamin D (10(-10), 10(-8), and 10(-6)?M). Gene and protein expression of hCAP-18 (p?<?0.05) showed an increase in sebocytes treated with vitamin D. Gene expression of hCAP-18 by treatment with vitamin D was blocked in sebocytes treated with VDR siRNA. In conclusion, treatment with vitamin D resulted in increased expression of LL-37 through the vitamin D receptor of cultured sebocytes.</P>
( Eugene Jeong ),( Ji Won Hong ),( Jung Ah Min ),( Dong Won Lee ),( Mi Yeung Sohn ),( Weon Ju Lee ),( Jun Young Lee ),( Young Min Park ) 대한피부과학회 2010 Annals of Dermatology Vol.23 No.1
Background: Although photodynamic therapy (PDT) is widely performed for acne, little is known about its exact therapeutic mechanism. Objective: We aimed to estimate the efficacy and safety of PDT on acne and to discover its mode of action. Methods: We performed PDT on 12 patients with mild to moderate acne. The clinical efficacy was assessed by counting the acne lesions and measuring the sebum secretion before and after PDT. In addition, we took biopsy samples from the peri-lesional areas before and after 3-session of PDT. To examine the degree of apoptosis of the sebaceous follicles, TUNEL assay was performed. To investigate the changes of toll-like receptor (TLR)-2 and TLR-4 expression after PDT, immunohistochemical stainings were also carried out. Finally, we performed TUNEL assay using the cultured sebocytes to confirm the apoptosis of sebocytes in vitro after PDT. Results: There was a significant reduction in the number of inflammatory acne lesions after PDT, compared to baseline (p<0.05). Sebum excretion significantly decreased 2 weeks after the first PDT session except for one patient (p<0.05). The TUNEL positive cells in the peri-lesional sebaceous glands after PDT markedly increased, compared with those of before PDT. A decrease in TLR-2 and TLR-4 expression by sebaceous glands and epidermis after PDT was 50% and 30%, respectively. Conclusion: Our results demonstrate that apoptosis of the sebaceous glands is associated with improvement of acne by PDT. PDT has shown to down-regulate TLR-2 and TLR-4 expression in the sebaceous glands and epidermis of acne patients. (Ann Dermatol 23(1) 23~32, 2011)
( Sang Lim Kim ),( Kyou Chae Lee ),( Jin Sub Lee ),( Yoon Hyuk Choi ),( Hyun Ho Son ),( Hong Dae Jung ),( Mi Yeung Sohn ),( Yong Hyun Jang ),( Seok Jong Lee ),( Do Won Kim ),( Weon Ju Lee ) 대한피부과학회 2013 대한피부과학회 학술발표대회집 Vol.65 No.2
Background: Acne is a inflammatory skin disorders caused by inflammatory biomarkers. Magnesium ascorbyl phosphate (MAP) is a stable precursor of vitamin C. It achieves a constant delivery of vitamin C into the skin and has antioxidative effects. Objectives: To evaluate changes in the expression of inflammatory biomarkers after treatment of cultured sebocytes with MAP Methods: RT-PCR, ELISA and western blotting were performed for inflammatory cytokines, matrix metalloproteinases (MMPs), antimicrobial peptides (AMPs) and keratinocyte differentiation markers before and after treatment of cultured sebocytes with MAP (10-2 mol), lipopolysaccharide (LPS) and a combination of MAP and LPS. RT-PCR and western blotting were also done for Toll-like receptor-4 (TLR-4) and NF-ĸB before and after treatment of cultured sebocytes with MAP, LPS and a combination of MAP and LPS. Results: The expressions of inflammatory cytokines (e.g., IL-1 β and IL-8) and MMPs (e.g., MMP-9) were decreased after treatment with MAP and a combination of MAP and LPS, and increased after treatment with LPS. AMPs (e.g., LL-37) were decreased after treatment with MAP and a combination of MAP and LPS, and increased after treatment with LPS. The expression of TLR-4 and NF-ĸB was decreased after treatment with MAP and a combination of MAP and LPS, and increased after treatment with LPS. Conclusion: MAP, which is a stable precursor of vitamin C, may be a good effective and alternative agent to improve inflammatory reaction in acne.
Vitamin D increases cathelicidin and decreases inflammatory cytokines in cultured sebocytes
( Jin Sub Lee ),( Sang Lim Kim ),( Hyun Wuk Cha ),( Chang Hyun Song ),( Kyung Duck Park ),( Hyun Jung Lim ),( Mi Yeung Sohn ),( Seok Jong Lee ),( Do Won Kim ),( Weon Ju Lee ) 대한피부과학회 2012 대한피부과학회 학술발표대회집 Vol.64 No.3