분리 세포의 단층세포 배양법에 의한 인체 비점막 상피세포의 배양

박호선,송시연,김용대,민명기,서장수,송계원 영남대학교의과대학 1998 Yeungnam University Journal of Medicine Vol.15 No.2

비중격 만곡증 환자의 비내수술시 체취한 정상하비갑개 점막조직으로부터 상피세포만을 분리세포의 단층배양법(monolayer culture of dissociated cells)으로 배양하여 비점막 상피세포 배양법을 정립하고 또한 배양된 세포가 상피세포임을 동정하기 위하여 간접 면역형광항체법으로 상피세포 특유의 cytokeratin을 확인하였고 투과전자현미경으로 상피세포만이 가지고 있는 교소체(desmosome) 및 장세사(tonofilament) 등을 확인하였으며 6회까지 계대배양을 할 수 있었다. 향후 본 연구에서 배양된 비점막 상피세포를 이용하여 알레르기성비염 등과 같은 비점막 염증성 질환의 병태생리에서 상피세포의 역할에 대한 연구와 호산구등 염증세포의 침윤과 상피세포의 손상에 접착분자 및 cytokine의 상호작용에 관한 연구가 계속될 수 있을 것으로 기대된다. Different techniques for culturing respiratory epithelial cells have been developed to overcome the limitations of studies on in vivo and on bioptic material. Traditionally, culture systems are divided into organ cultures, explant cultures and dissociated cell cultures. The first two contain both epithelial and non-epithelial cells. However, in monolayer cultures of dissociated cells only epithelial cells are present, the effects observed are caused by a pure epithelial responses. The purpose of this study is to establish primary culture method of human nasal epithelium (HNEC) by monolayer culture of dissociated cells to evaluate the role of the epithelial cells in the allergic and non-allergic nasal inflammatory reactions. HNEC was prepared by primary culture method of monolayer culture of dissociated cells from human inferior nasal turbinate mucosa of septal deviation patients. Primary cultured cells were characterized by indirect immunofluorescence assay and transmission electron microscopy. The immunoreactivities of cytokeratin-pan and cytokeratin No. 8 were observed in cultured HNEC. However, the immoreactivities of vimentin and von Willebrand factor were not observed in cultured HNEC. The tonofilaments and desmosome were observed in cultured HNCE. The cultured epithelial cells were identified to be pure nasal epithelial cells. The monolayer culture of dissociated cells could successfully be employed for further study to investigate the role of the epithelial cells in allergic or non-allergic nasal inflammatory diseases.

화학적 폐 손상시 상피세포 장벽의 특성

서덕준,김준연 동아대학교 의과대학 부설 산업의학연구소 1992 산업의학연구소 논총 Vol.- No.1

폐 상피세포 장벽은 다른 tight한 생체세포의 장벽에서와 마찬가지로 높은 조직저항을 보이며, 따라서 용매와 용질의 이동을 극히 제한한다. 이러한 수동적인 장벽의 특성은 폐의 여러조건에 의하여 심하게 변할 수 있으며 요즈음 폐 상피세포 손상의 장애시 그 기전이 활발하게 연구되고 있다. 정상적인 tight 한 폐 상피세포 장벽은 간질액과 혈관내로 부터 용질이 빠져 나오는 것을 막음으로써 폐의 일차적인 기본 기능인 기체교환을 하는데 있어 정상적으로 폐표면을 항상 건조하게 유지시킬 수 있는 것이다. 그러나 현재까지의 결과는 거의 모두가 작은실험 동물들의 전체 폐를 이용한 것으로 그 기전을 알기가 어렵고, 인체의 폐 상피세포에서의 물질이동에 대한 특성은 거의 알려진 바가 없다. 화학물질에 의한 인체 폐 부종의 기전을 알기 위하여 여러 실험 모델에서 얻은 결과를 통하여 추후 진단 및 그 자료 방침을 정할 수 있게 될 것으로 생각된다. The alveolar epithelial barrier permits only restricted passage of solutes examplified by a high resistance and equivalent pore properties similar to those of other tight biological barriers. These passive barrier properties may be markedly altered after insults to the lung, perhaps providing insights into the mechanisms of alveolar epithelial injury. The normally tight alveolar epithelium serves as an effective barrier against the leak of solutes from interstitial and vascular spaces into alveolar spaces, thereby helping to maintain the normal "dry" environment required for efficient gas exchange. Until recently, the majority of investigations of alveolar fluid balance relied on intact lung models. The information obtained from these intact mammalian lung studies in vivo and in vitro, if useful, can be difficult to interpret mechanistically, largely because of the anatomical complexity of the lung. For more precise information about alveolar epithelial properties, studies of isolated preparations have been done, providing useful additional approaches to underlying transport mechanisms and pathways in the alveolar-capillary wall. In this review article, I introduced the methods being used for study of barrier function in addition to intact lung investigation. And I discussed the barrier properties of the alveolar epithelium relevant to alveolar fluid balance in chemically injured lung, citing the pertinent supporting experimental data where appropriate.

Physical stress regulates the expansion patterns of epithelial monolayer

Youngbin Cho,Bomi Gweon,Jacob Notbohm,Dayoung Kim,Hyuntae Jeong,Hwanseok Jang,David Helfman,Yongdoo Park,Jeffrey Fredberg,Jennifer H. Shin 대한기계학회 2019 대한기계학회 춘추학술대회 Vol.2019 No.4

In Vitro Maturation of Round Spermatids Using Porcine Oviduct Epithelial Cell Monolayer Condition Medium

Jabed Md. Anower,Kamal Tania,Lee Seung-Min,Kim Byung Ki The Korean Society of Animal Reproduction 2005 Reproductive & developmental biology Vol.29 No.4

Porcine oviduct epithelial cells (POEC) are widely used in co-culture experiments to improve early embryonic development, in vitro fertilization in embryo transfer programs for domestic animals and in vitro maturation of immature germ cells. POEC were mechanically isolated and cultured in tissue culture medium 199. Cells grew continuously, and confluent monolayers were formed after 7 days. After forming confluent monolayer of epithelial cells, supernatant was collected as the condition medium for maturing round spermatids in vitro. Round spermatids were also separated mechanically and cultured in the POEC condition medium. In this study we observed that $20\%$ of round spermatid cultured were matured into elongating spermatid after 24 h, and about $10\%$ of round spermatid cultured showed complete elongation (elongated spermatid) within $24\~48$ h of in vitro culture. No further development was observed within $50\~72$ h and transformed cells lost their viability after 72 h. These preliminary findings suggest that the condition medium from POEC may be possible to overcome the round spermatid block by improving the milieu of culture system.

화학적 폐 손상시 폐 상피세포 장벽의 특성

서덕준,김준연 大韓産業醫學會 1991 대한직업환경의학회지 Vol.3 No.2

The alveolar epithelial barrier permits only restricted passage of solutes, exemplified by a high resistance and equivalent pore properties similar to those of other tight biological barriers. These passive barrier properties may be markedly altered after insults to the lung, perhaps providing insights into the mechanisms of alveolar epithelial injury. The normally tight alveolar epithelium serves as an effective barrier against the leak of solutes from interstitial and vascular spaces into alveolar spaces, thereby helping to maintain the normal "dry" environment required for efficient gas exchange. Until recently, the majority of investigations of alveolar fluid balance relied on intact lung models. The information obtained from these intact mammalian lung studies in vivo and in vitro, if useful, can be difficult to interpret mechanistically, largely because of the anatomical complexity of the lung. For more precise information about alveolar epithelial properties, studies of isolated preparations have been done, providing useful additional approaches to underlying transport machanisms and pathways in the alveolar-capillary wall. In this review article, I introduced the methods being used for study of barrier function in addition to intact lung investigation. And I discussed the barrier properties of the alveolar epithelium relevant to alveolar fluid balance in chemically injured lung, citing the pertinent supporting experimental data where appropriate.

물리적 힘 가시화를 통한 세포 군집의 확장 패턴 분석

조영빈(Youngbin Cho),신현정(Jennifer H. Shin),권보미(Bomi Gweon),Jacob Notbohm,김다영(Dayoung Kim),정현태(Hyuntae Jeong),장환석(Hwanseok Jang),David Helfman,박용두(Yongdoo Park),Jeffrey Fredberg 한국가시화정보학회 2018 한국가시화정보학회 학술발표대회 논문집 Vol.2018 No.11

In Vitro Maturation of Round Spermatids Using Porcine Oviduct Epithelial Cell Monolayer Condition Medium

Md. Anower Jabed,Tania Kamal,Seung-Min Lee,Byung Ki Kim 한국동물생명공학회(구 한국동물번식학회) 2005 Reproductive & developmental biology Vol.29 No.4

Porcine oviduct epithelial cells (POEC) are widely used in co-culture experiments to improve early embryonic development, in vitro fertilization in embryo transfer programs for domestic animals and in vitro maturation of immature germ cells. POEC were mechanically isolated and cultured in tissue culture medium 199. Cells grew continuously, and confluent monolayers were formed after 7 days. After forming confluent monolayer of epithelial cells, supernatant was collected as the condition medium for maturing round spermatids in vitro. Round spermatids were also separated mechanically and cultured in the POEC condition medium. In this study we observed that 20% of round spermatid cultured were matured into elongating spermatid after 24 h, and about 10% of round spermatid cultured showed complete elongation (elongated spermatid) within 24~48 h of in vitro culture. No further development was observed within 50~72 h and transformed cells lost their viability after 72 h. These preliminary findings suggest that the condition medium from POEC may be possible to overcome the round spermatid block by improving the milieu of culture system.

In vitro Nasal Cell Culture Systems for Drug Transport Studies

조현종,Ubonvan Termsarasab,김정선,김대덕 한국약제학회 2010 Journal of Pharmaceutical Investigation Vol.40 No.6

Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the “passage” culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

In vitro Nasal cell Culture Systems for Drug Transport Studies

( Hyun Jong Cho ),( Ubonvan Termsarsab ),( Jung Sun Kim ),( Dae Duk Kim ) 한국약제학회 2010 Journal of Pharmaceutical Investigation Vol.40 No.6

Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the “passage” culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

In vitro Nasal Cell Culture Systems for Drug Transport Studies

Cho, Hyun-Jong,Termsarasab, Ubonvan,Kim, Jung-Sun,Kim, Dae-Duk The Korean Society of Pharmaceutical Sciences and 2010 Journal of Pharmaceutical Investigation Vol.40 No.6

Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.