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황정욱 ( Jung Wook Hwang ),이희삼 ( Heui Sam Lee ),김호진 ( Ho Jin Kim ),김규오 ( Kyu Oh Kim ),최용수 ( Yong Soo Choi ) 한국잠사학회 2012 한국잠사곤충학회지 Vol.50 No.2
Silk protein has been explored to be used for biomedical applications for several decades. However, it has not been used in this field cause to their irreversible crystallization after dissolving in water. The existing methods of silk protein hydrolysis using silkworm cocoon were used with harmful solvents and through a very complicated process. Therefore, we have developed novel methods for the production of water-soluble hydrolysate using silkworm gland. We manufactured two types of silkworm gland-derived hydrolysate (water-soluble SGH, SSGH; total SGH, TSGH) and compared the characteristics with commercial cocoon-derived sericin hydrolysate (CSH). The molecular weight of SGH ranged from 7 to 50 kDa (SSGH) and 5 to 15 kDa (TSGH) within glycine, alanine, and aspartic acid as a main amino acid composition. In contrast, CSH ranged from 15 to 50 kDa within serine and aspartic acid. The results of FTIR implied that SGH was more soluble form than CSH, as shown by the decrease in the β-sheet structure at 1630 cm-1 on amide I peak. In comparison with 10% fetal bovine serum, 0.1% (1 mg/ml) SSGH had equivalent effect on the proliferation of human dermal fibroblasts and mesenchymal stem cells. All results of the SSGH made by novel manufacturing process indicate the SSGH is more preferable as a culture medium supplement than cocoon-derived sericin.
정지혜(Ji-Hye Jung),황정욱(Jung Wook Hwang),김호진(Hojin Kim),차현명(Hyun-Myoung Cha),김동일(Dong-Il Kim),최용수(Yong-Soo Choi) 한국생물공학회 2013 KSBB Journal Vol.28 No.3
Silkworm-derived silk materials which contain 45 to 55% protein (18 amino acids, including 8 essential amino acids) have free radical-scavenging activities. In the present study, we investigated the whitening and anti-aging effects of silkworm gland under various conditions with B16F1 melanoma cells and human dermal fibroblasts. To evaluate the toxicity of soluble gland hydrolysate (SGH), mouse-derived B16F1 melanoma cells were treated with 0.2~5 mg/mL, cytotoxicity was not observed at all concentrations. The tyrosinase and elastase activities were significantly decreased as dosage levels of SGH increased. In addition, cell death was decreased by treated with SGH and antioxidative activity was the most effective in the SGH treatment. These findings suggest that SGH may be used as a potential functional biomaterial in having the skin whitening effect and anti-aging activity by inhibition of tyrosinase and elastase activities.
형질전환 Chinese Hamster Ovary 세포에서 Albumin-erythropoietin의 생산시 Silkworm Gland Hydrolysate의 효과
최민호(Min-Ho Choi),차현명(Hyun-Myoung Cha),김선미(Sun-Mi Kim),최용수(Yong-Soo Choi),김동일(Dong-Il Kim) 한국생물공학회 2013 KSBB Journal Vol.28 No.2
To date, various strategies have been studied to increase specific productivity in Chinese hamster ovary (CHO) cell cultures. Also, albumin-fusion platform is being applied to other important bioactive peptides with short half-lives. Here, we investigated the effects of silkworm gland hydrolysate (SGH) on the production of albumin-erythropoietin (Alb-EPO) in transgenic CHO cells. The viable cell density of CHO cells was increased by 13% in the medium containing 1 mg/mL SGH higher than in the control medium without SGH. In addition, the production of Alb-EPO was also 1.26-fold enhanced by reducing the early apoptosis of CHO cells. In conclusion, SGH could be used as a useful supplement for the enhancement of recombinant protein production.
중간엽줄기세포 증식을 위한 누에 실샘 유래 무혈청 배지 첨가제
차현명 ( Hyun Myoung Cha ),김선미 ( Sun Mi Kim ),최용수 ( Yong Soo Choi ),박지성 ( Ji Sung Park ),임진혁 ( Jin Hyeok Lim ),황슬기 ( Seul Gee Hwang ),김동일 ( Dong Il Kim ) 한국조직공학과 재생의학회 2015 조직공학과 재생의학 Vol.12 No.1s
Mesenchymal stem cells (MSCs) are must be cultured in chemically defined, xeno-free culture system for cell-based therapies. One major overcome for their clinical use is the unsafety of fetal bovine serum (FBS), which is a crucial part of all media currently used for the culture of MSCs. Previously, we developed novel methods the production of silkworm gland hydrolysate (SGH). Also, we showed the effect of the SGH on the cell growth promotion. We demonstrated whether the characteristics of MSCs were maintained in the serum-free media containing SGH during long-term cultivation. The results of microscopic observation showed that the morphology of stem cells were not different between the media containing 10% FBS and 1 mg/mL SGH. After the long-term culture of MSCs in serum-free media containing SGH, the rate of MSCs growth was well maintained. The analysis of anti-apoptotic effect of SGH showed that early apoptosis of MSCs was reduced by 3.7-fold. Furthermore, MSCs specific surface markers and stemness gene expression (Nanog, Sox2) were preserved as that of MSCs in the media containing 10% FBS. In conclusion, we demonstrate that the SGH can be used as useful supplement for MSCs cultivation in serumfree media.