냉동 보관된 백서태자 중뇌 신경세포의 생존율과 신경돌기 형성율에 대한 연구

이 언,김영보,유영미 대한신경외과학회 1999 Journal of Korean neurosurgical society Vol.28 No.2

동종동맥판 혈관내피세포의 생육성 평가에 관한 연구

임창영,홍은경 大韓胸部外科學會 1997 Journal of Chest Surgery (J Chest Surg) Vol.30 No.6

A Rapid and Simple flow Cytometric Method for Measuring Cell Viability Using Propidium Iodide Staining and Forward Scatter Measurement

Lee, Yong Suk,Youn, Sang Woong,Kim, Kyu Han,Park, Kyoung Chan Korean Dermatological Association 1996 Annals of Dermatology Vol.8 No.3

Photoacoustic Monitoring of the Viability of Mesenchymal Stem Cells Labeled with Indocyanine Green

Yoo, J.M.,Yun, C.,Bui, N.Q.,Oh, J.,Nam, S.Y. Elsevier 2019 IRBM Vol.40 No.1

<P><B>Abstract</B></P> <P><B>Background</B></P> <P>Stem cell therapy has a huge potential to enhance the recovery of damaged tissues and organs. However, it has been reported that majority of implanted stem cells cannot survive after implantation. Therefore, noninvasive monitoring of stem cell viability is essential to estimate the efficacy of stem cell therapy. However, current imaging methods have disadvantages for monitoring of stem cell viability such as cost, penetration depth, and safety. To overcome the limitations, photoacoustic imaging well known for sufficient penetration depth, relatively low cost, and non-ionizing radiation can be a novel alternative assessment method of stem cell viability.</P> <P><B>Methods</B></P> <P>In this study, indocyanine green was used as exogenous photoacoustic contrast agents to label mesenchymal stem cells. The photoacoustic signals were acquired before and after the cell death and quantified to monitor photoacoustic signal changes related to the cell viability.</P> <P><B>Results</B></P> <P>The fluorescence intensity changes of ICG labeled MSCs corresponded to decrease of PA intensity after cell death. Furthermore, the PA imaging of MSCs showed similarity between the PA intensity and the cell viability.</P> <P><B>Conclusion</B></P> <P>The experimental results imply the feasibility of noninvasive detection of stem cell viability during therapeutic procedures.</P> <P><B>Highlights</B></P> <P> <UL> <LI> ICG can be used as an MSC labeling probe for PA imaging. </LI> <LI> PA imaging of MSCs showed similarity between the PA intensity and the cell viability. </LI> <LI> PA imaging with ICG labeling is an alternative to detect MSCs viability non-invasively. </LI> </UL> </P>

Phosphate-Induced Rat Vascular Smooth Muscle Cell Calcification and the Implication of Zinc Deficiency in A7r5 Cell Viability

Mee-Young Shin,In-Sook Kwun 한국식품영양과학회 2013 Preventive Nutrition and Food Science Vol.18 No.2

The calcification of vascular smooth muscle cells (VSMCs) is considered one of the major contributors for vascular disease. Phosphate is known as the inducer for VSMC calcification. In this study, we assessed whether phosphate affected cell viability and fetuin-A, a calcification inhibitor protein, both which are related to VSMC calcification. Also, VSMC viability by zinc level was assessed. The results showed that phosphate increased Ca and P deposition in VSMCs (A7r5 cell line, rat aorta origin). This phosphate-induced Ca and P deposition was consistent with the decreased A7r5 cell viability (P<0.05), which implies phosphate-induced calcification in A7r5 cells might be due to the decreased VSMC cell viability. As phosphate increased, the protein expression of fetuin-A protein was up-regulated. A7r5 cell viability decreased as the addition of cellular zinc level was decreased (P<0.05). The results suggested that zinc deficiency causes the decreased cell viability and it would be the future study to clarify how zinc does act for VSMC cell viability. The results suggest that the decreased VSMC viability by high P or low Zn in VSMCs may be the risk factor for vascular disease.

Phosphate-Induced Rat Vascular Smooth Muscle Cell Calcification and the Implication of Zinc Deficiency in A7r5 Cell Viability

Shin, Mee-Young,Kwun, In-Sook The Korean Society of Food Science and Nutrition 2013 Preventive Nutrition and Food Science Vol.18 No.2

The calcification of vascular smooth muscle cells (VSMCs) is considered one of the major contributors for vascular disease. Phosphate is known as the inducer for VSMC calcification. In this study, we assessed whether phosphate affected cell viability and fetuin-A, a calcification inhibitor protein, both which are related to VSMC calcification. Also, VSMC viability by zinc level was assessed. The results showed that phosphate increased Ca and P deposition in VSMCs (A7r5 cell line, rat aorta origin). This phosphate-induced Ca and P deposition was consistent with the decreased A7r5 cell viability (P<0.05), which implies phosphate-induced calcification in A7r5 cells might be due to the decreased VSMC cell viability. As phosphate increased, the protein expression of fetuin-A protein was up-regulated. A7r5 cell viability decreased as the addition of cellular zinc level was decreased (P<0.05). The results suggested that zinc deficiency causes the decreased cell viability and it would be the future study to clarify how zinc does act for VSMC cell viability. The results suggest that the decreased VSMC viability by high P or low Zn in VSMCs may be the risk factor for vascular disease.

Phosphate-Induced Rat Vascular Smooth Muscle Cell Calcification and the Implication of Zinc Deficiency in A7r5 Cell Viability

신미영,권인숙 한국식품영양과학회 2013 Preventive Nutrition and Food Science Vol.18 No.2

The calcification of vascular smooth muscle cells (VSMCs) is considered one of the major contributors for vascular disease. Phosphate is known as the inducer for VSMC calcification. In this study, we assessed whether phosphate affected cell viability and fetuin-A, a calcification inhibitor protein, both which are related to VSMC calcification. Also, VSMC viability by zinc level was assessed. The results showed that phosphate increased Ca and P deposition in VSMCs (A7r5 cell line, rat aorta origin). This phosphate-induced Ca and P deposition was consistent with the decreased A7r5 cell viability (P<0.05), which implies phosphate-induced calcification in A7r5 cells might be due to the decreased VSMC cell viability. As phosphate increased, the protein expression of fetuin-A protein was up-regulated. A7r5 cell viability decreased as the addition of cellular zinc level was decreased (P<0.05). The results suggested that zinc deficiency causes the decreased cell viability and it would be the future study to clarify how zinc does act for VSMC cell viability. The results suggest that the decreased VSMC viability by high P or low Zn in VSMCs may be the risk factor for vascular disease.

온혈허혈시간과 냉동보존온도와 보존액 조성에 따른 기관의 생육성 비교

사영조,박재길,심성보,진웅,문영규,이선희,조건현 대한흉부외과학회 2009 Journal of Chest Surgery (J Chest Surg) Vol.42 No.3

Background: Tracheal reconstruction after extended tracheal resection still remains as a major surgical challenge because good clinical outcomes are usually correlated with limited tracheal resection. Recent investigations with a using cryopreserved trachea for the reconstruction of a trachea have been carried out to overcome this problem. In this study, we analyzed viability of tracheas, which is an important determining factor for the success of transplanting a cryopreserved trachea and the development of post-transplantation tracheal stenosis, according to three different experimental factors: 1) the warm-ischemic time, 2) the cryopreservation solution and 3) the preserving temperature, to determine a better cryopreservation protocol and a better composition of the cryopreservation solution. Material and Method: Rats tracheas were harvested for different warm-ischemic times (0 hr, 12 hrs, 24 hrs). The tracheas were treated with recombinant insulin growth factor-1 (IGF-1) and they were stored at three different temperatures (4℃, −80℃, −196℃) for two weeks. After two weeks, we thawed the stored trachea and isolated the cells of the tracheas with using type II collagenase. We cultured the cells for seven days and then we compared the cellular viability by the MTT reduction assay. Result: Though cryopreservation is required to preserve a trachea for a longer time period, the viability of the tracheas stored at −80℃ and −196℃ was significantly reduced compared to that of the tracheas stored at 4℃. The viability of the tracheas with warm-ischemic times of 12 hrs and 24 hrs was also reduced in comparison to the tracheas with a warm-ischemic time of 0 hrs. Our data showed that the warm ischemic time and the parameters of cryopreservation negatively affect on trachea viability. However, a cryopresrvation solution containing IGF-1 improved the cellular viability better than the existing cryopreservation solution. For the warm ischemic time group of 0 hr, the addition of IGF-1 improved the viability of trachea at all the preserving temperatures. Conclusion: These experiments demonstrate that the viability of a cryopreserved trachea can be improved by modifying the components of the cryopreservation solution with the addition of IGF-1 and reducing the warm-ischemic time. 배경: 기관재건술은 제한된 경우에서만 뚜렷한 효과를 얻을 수 있어 광범위한 기관절제술 후의 기관재건술은 아직 의학적으로 해결되지 못하고 있는 난제들 중의 하나로 남아 있다. 이 어려운 문제를 해결하기 위한 방법으로 냉동 보존된 기관을 이용하여 기관을 재건하려는 노력이 이루어지고 있다. 냉동 보존된 기관을 이용한 재건에서는 수술의 성공 여부에 가장 중요한 결정인자가 바로 기관의 생육성이다. 이에 저자들은 기관의 냉동 보존 시 냉동 보존액의 조성과 온혈허혈시간의 정도에 따른 차이, 그리고 보존온도의 정도에 따른 차이에 따른 기관연골의 생육성의 차이를 비교 검토하여, 보다 나은 냉동 보존방법을 알아보고자 하였다. 대상 및 방법: 정해진 온혈허혈시간(0시간, 12시간, 24시간)의 경과 후 쥐의 기관을 채취하여, recombinant insulin growth factor-1 (IGF-1)을 처치하고 3가지의 보존 온도(4℃, −80℃, −196℃)에서 2주간 보존하였다. 보존 후 해동하여 type II collagenase효소를 이용하여 기관의 세포를 채취하였다. 채취한 세포를 7일간 배양한 뒤 MTT reduction assay를 이용하여 각 군의 기관 세포의 생육성을 비교하였다. 결과: 기관을 오랜 기간 보존하기 위해서는 냉동보존은 필요하지만, −80℃와 −196℃에서의 냉동 보존은 대조군과 4℃ 보존군에 비해 통계적으로 유의할 정도로 기관의 생육성을 감소시키는 것으로 관찰되었고, 12시간과 24시간의 온혈허혈시간도 온혈허혈시간 0시간 군에 비해 기관의 생육성을 감소시키는 것으로 관찰되었다. IGF-1을 첨가한 냉동 보존액은 기존의 냉동 보존액보다 기관의 생육성을 향상시키는 것을 확인할 수 있었고, IGF-1로 인한 생육성 향상은 4℃ 보존군에서는 모든 온혈허혈시간에서, 온혈허혈시간 0시간 군에서는 모든 보존온도에서 관찰되었다. 결론: 온혈허혈시간을 최대한 줄이며 냉동 보존액에 IGF-1을 첨가하여 보존액의 조성을 조정함으로써, 냉동 보존 시 보다 나은 기관의 생육성을 유지할 수 있을 것으로 판단되었다.

Human Neutrophil의 수명연장과 Superoxide 생산에 관여하는 미지의 Monocyte 생성물질

허억,배진우 大韓免疫學會 1993 大韓免疫學會誌 Vol.15 No.-

It has long been known that neutrophils are quickly infilterated and recruited to infected sites and then kill invaders by phagocytic action. Unfortunatly it is not yet revealed which molecule or cytokine is involved in the phagocytic action and viability sustaining activity of neutrophils. The aim of this study was whether lipopolysaccharide (LPS)-stimulated monocyte may control those neutrophil actions. Human peripheral blood monocytes and neutrophils were isolated by Ficollpaque density sedimentation from heparin anti-coagulated blood of healthy adult donors. After preparation of these cells, the purity of both was more than 90%. Monocytes stimulated in various dose(0.1-10pug/ml) of LPS for various times of incubation(0-3 days). and then LPS-stimulated monocyte conditioned medium was collected in order to find an optimal dose and incubation time for the neutrophil viability. It was found out that 3,ug/ml of LPS in 24 hours incubation was maximal effective condition for the activity of neutrophil sustaining viability. Monocyte conditioned medium (MCM) under this condition was used for the comparison with LPS-nonstimulated monocyte conditioned medium or enriched medium alone. When neutrophils were stimulated with each medium for 1-3 days, the activity of neutrophil sustaining viability with MCM was significantly higher than the activity with other medium (in 1 day of culture, 72-1:8 vs 4311:7 vs 17 ±10; p <O. 01). The superoxide production of neutrophil stimulated with MCM for 24 hours incubation was significantly higher than that with other medium under fMLP doses of 0.1-100,uM (p <0.01). Under fMLP l,uM, the superoxide production is predominantly different between them(23.8±2 vs 10.3±3 vs 7.8±1.6). The maximal effective dose of GM-CSF(granulocyte/macrophage colony stimulating factor ; 10pM) enhanced the neutrophil viability in 1 day of culture (50 ± 4%) . In the study to assess whether MCM contains GM-CSF, anti-GM-CSF antibody slightly blocked the MCM-dependent neutrophil viability(73±9 vs 50±12; p <0.07), indicating that MCM might not contain GM-CSF. These data indicate that LPS-stimulated monocyte surely product a factor for the neutrophil sustaining viability and the enhancement of superoxide production, suggesting that a factor is not GMCSF. If more than one factor were producted form LPS-stimulated monocyte, one minor factor might be GM-CSF.

Label-free Measurement of Cell Viability via Counting Cells Attached on Affinity Substrates

Ahn, Junhyoung,Park, Jina,Kim, Yeon-Gu,Lee, Eun Gyo,Kim, Min-Gon,Shin, Yong-Beom 한국생물공학회 2014 Biotechnology and Bioprocess Engineering Vol.19 No.2

The commonly used trypan blue dye exclusion method and other modified cell viability methods, such as fluorescein dye and tetrazolium dye exclusion, artificially introduce toxic chemicals to cells and, thus, alter cellular organelles when measuring cell viability. Therefore, cell viability could be affected by the processes currently used to observe viability. In this study, the cell viability of Chinese hamster ovary (CHO) cells was measured by simply counting attached cells after the cultured CHO cells were attached on a Concanavalin A (Con A) substrate. The efficiency of cell attachment to Con A surfaces was different for live and dead cells allowing the cell viability of CHO cells to be measured without any chemical modifications to the cells.