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박정극,김영진,송계용,유보영,Youn-Ho Shin,윤희훈,Sung-Joo Hwang 한국생물공학회 2007 Biotechnology and Bioprocess Engineering Vol.12 No.1
Hair follicle is a small but very complex and dynamic miniorgan of the human body. It is easy to isolate and culture mesenchymal cells but not epithelial cells of hair follicle. It is necessary for intact and healthy outer root sheath (ORS) cells to be isolated and cultured. In this study we developed an appropriate isolation method to yield 6.4 ± 0.75 × 104 cells/hair follicle, which is about 9-fold comparing to our previous data. This yield was achieved by modifications such as different kinds of enzyme uses, fragmentation, and mechanical stimuli. Especially we detected that the different kinds of isolation enzyme could affect proliferation of ORS cells during primary culture. In addition, bovine pituitary extract (BPE) was needed for ORS cells to proliferate and to form colonies under serum-free, feeder layer-free culture condition, but type I collagen as a substratum did not have any positive effect. Moreover, ORS cells under BPE-added condition contained stem/progenitor cells expressing β1-integrin, CK19, and CD34. These results can provide useful cell culture information, not only in the study of hair biology but also in the field of tissue engineering and cell therapy for the treatment of alopecia.
박정극,송계용,권순용,Kyung-Min Choi,Young-Kwon Seo,윤희훈,Hwa-Sung Lee 한국생물공학회 2007 Biotechnology and Bioprocess Engineering Vol.12 No.6
To support and enhance the in vitro growth and activity of mesenchymal stem cells (MSCs), the cell culture medium may be supplemented with various proteins and factors to mimic the physiological environment in which the cells optimally proliferate and differentiate. In this study, the effects of mechanical factors on cellular metabolic responses were investigated experimentally using a bioreactor. The effects of various chemical factors, such as growth factors, cytokines, and hormones, were also investigated. Based on previous reports demonstrating the important roles of mechanical factors in the growth and activity of MSCs, we sought to evaluate the effects of mechanical stimuli on the proliferation of bone marrow-derived MSCs using a cell training bioreactor that imposed cyclic mechanical stretch, with parameters of 240 min/day, 0.03 Hz, and 5~15% strain. The application of cyclic stretch (5~15% strain) to the MSCs enhanced their proliferation during the early stage (3 days), but not the late stage (14 days), of batch culture. Mechanical stretch did not increase the release of lactate dehydrogenase (LDH) from the MSCs during culture. Appropriate levels of mechanical stretch (5~10% strain) increased collagen synthesis, but did not alter MSC surface antigen expression. It is thought that the appropriate level of mechanical stretch was able to serve as a potent positive modulator of MSC proliferation during the initial stages of culture.
FTIR을 이용한 상용 흡착제의 바이오가스 실록산 흡착 특성에 관한 실험적 연구
박정극,허광범,이정근,이남훈 한국폐기물자원순환학회 2018 한국폐기물자원순환학회지 Vol.35 No.6
Removal of siloxane compounds is very important to protecting the biogas energy conversion system from decreased efficiency and parts damage. Among various siloxane removal technologies, adsorption towers are mostly used for performance and ease of operation. However, due to the difficulty of measuring the concentration of siloxane compounds in the gas stream and the complicated matrix of siloxane compounds, adsorption characteristics are not well known. In this study, the adsorption characteristics for multi siloxane components are experimentally studied. Four siloxane components are vaporized in the nitrogen stream supplied continuously to a lab-scale adsorption tank with commercially available silica gel or activated carbon and an FTIR (Fourier-transform infrared spectroscopy) analyzer was used for the online siloxane analysis to find out the adsorption characteristics. While a mixture of L2, L5, D4 and D5 adsorption capacity of silica gel and activated carbon are similar -11.13 and 11.56wt% respectively-adsorption characteristics of each adsorbent was well distinguished in terms of breakthrough behavior. Silica gel shows sequential breakthrough for each siloxane compound and a more noticeable unique time range for Rc > 1, while relatively simultaneous breakthrough was shown for activated carbon adsorbents.
박정극,송계용,김기호,이희구,양은경,안재일,장인근,이두훈,Young-Kwon Seo,Hee-Hoon Yoon,Youn-Ho Shin,Jae-Chan Kim 한국생물공학회 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3
Many researchers have employed cryopreserved amniotic membrane (CAM) in the treatment of a severely damaged cornea, using corneal epithelial cells cultured on an amniotic membrane (AM). In this study, two Teflon rings were made for culturing the cells on the LAM and CAM, and were then used to support the AM, which is referred to in this paper as an Ahns AM supporter. The primary corneal epithelial cells were obtained from the limbus, using an explantation method. The corneal epithelium could be reconstructed by culturing the third-passage corneal epithelial cells on the AM. A lyophilized amniotic membrane (LAM) has a higher rate of graft take, a longer shelf life, is easier to store, and safer, due to gamma irradiation, than a CAM. The corneal epithelium reconstructed on the LAM and CAM, supported by the two-Teflon rings, was similar to normal corneal epithelium. However, the advantages of the LAM over that of the CAM make the former more useful. The reconstruction model of the corneal epithelium, using AM, is considered as a good in vitro model for transplantation of cornel epithelium into patients with a severely damaged cornea.
박정극,김영진,양은경,장이섭,유보영,서영권,이두훈,신윤호,이경미,송계용,서성준,왕성주,박창서 한국생물공학회 2003 Biotechnology and Bioprocess Engineering Vol.8 No.2
Obtaining a sufficient amount of healthy keratinocytes from a small tissue is difficult. However, ORS cells can be a good source of epithelium since they are easily obtainable and patients do not have to suffer from scar formation at donor sites. Accordingly, the current study modified the conventional primary culture technique to overcome the low propagation and easy aging of epithelial cells during culturing. In a conventional primary culture, the average yield of human ORS cells is 2.1 x 103 cells/follicle based on direct incubation in a trypsin (0.1%)/EDTA (0.02%) solution for 15 min at 37oC, however, our modified method was able to obtain about 6.9 x 103 cells/follicle using a two-step enzyme digestion method involving dispase (1.2 U/mL) and a trypsin (0.1%)/EDTA (0.02%) solution. Thus, the yield of primary cultured ORS cells could be increasd three times higher. Furthermore, a total of 2.0 x 107 cells was obtained in a serum-free medium, while a modified E-medium with mitomycin C-treated feeder cells produced a total of 6.3 x 107 cells over 17 days when starting with 7.5 x 104 cells. Finally, we confirmed the effectiveness of our ORS cell isolation method by presenting their ability for reconstructing the bioartificial skin epithelium in vitro.