The inflammatory responses are complicated events that involve many immune cells from resident tissue and infiltrated cells from blood vessel. In skin and cornea, epidermal lining provides a first line of defense against microbial pathogens as well as physical and chemical insult. To evaluate cosmetic ingredients for inflammation and allergy, a hypersensitivity of immune reaction, we conventionally utilized either in vivo animal models or in vitro immune cells. In this study, we developed a new alternative method for inflammation and allergy testing using co-culture of 3D- reconstructed skin and cornea with monitoring immune cells which represents in vitro substitute for functional epidermal barrier and multiple immune responses in the tissue. Firstly, we generated 3D- reconstructed skin and cornea with cells from human skin and rabbit cornea using transwell. Specific markers for basal and differentiated layer in normal skin or cornea, either K5 and K10 or K5 and K3+12, were well expressed in 3D skin or cornea, respectively. We applied RAW264.7 and RBL-2H3 as monitoring immune cells for inflammatory and allergic responses in co-culture system. LPS-induced immune responses in RAW 264.7 cells were examined. Decreased expression levels of IL-6, TNF-alpha, IL-10, iNOS, and COX-2 were observed in co-culture system compared to those of the direct administration in RAW 264.7 cells. In addition, DNP-IgE-mediated allergic responses were attenuated in co-culture system which was monitored by b-hexosaminisases activity in RBL-2H3. These data suggests that epidermal barriers of 3D-reconstruced skin and cornea provide a protective role under inflammatory environment in a similar mechanism as with intact in vivo condition. Especially, RAW 264.7 cells played dual roles mediating inflammatory responses and acted as anti- inflammatory dermal macrophages in the skin. These results suggest that this in vitro co-culture model system can be used as a novel alternative method for testing immune responses in the skin and cornea.
저자
Eunkyung CHUNG [ Department of Genetic Engineering, College of Life Science and Graduate School of Biotechnology, Kyung Hee University, Seocheon-dong, Kiheung-gu, Yong In, 446-701, Korea. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
이 법인은 생물 공학의 발전과 보급에 이바지하고, 회원 상호 간의 연구 협력과 친목을 도모함을 목적으로 한다
1. 생물공학 분야의 발전을 위한 연구 협력
2. 생물공학의 실용화를 촉진시키기 위한 산학 협동
3. 학술연구 발표회, 강연회, 연수회 등 학술활동의 개최
4. 국,영문 학술지,소식지,학술회의 Proceedings 및 학술도서의 발간
5. 생물공학 발전을 위한 정책 건의
6. 기타 국제 교류 등 본 학회의 목적 달성을 위한 제반 활동