이 작품은 콜럼버스 장 9671 (윈저, 온타리오), 그리고 CIHR 프레더릭 들소와 데니스 엄마에게 수여 찰스 베스트 캐나다 대학원 장학금의 기사에 의해 지원되었습니다. 촬영된 현미경으로 그들의 도움 로버트 호지와 엘리자베스 Fidalgo 다 실바에게 감사합니다. 촬영된 현미경 비디오를 편집 케이티 Facecchia에 감사합니다. 우리는 또한이 원고의 비판적 검토를 위해 Sudipa 6월 Chatterjee와 필립 Tremblay 감사드립니다. 이 작품은 2010 년 암 상대로 자신의 연이은 승전 케빈 Couvillon의 메모리에 전념하고 있습니다.

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관심의 어떠한 충돌 선언 없습니다.

PST 및 유사 화합물은 안티 - 암 속성에게 11-15,21을 가지고 보여왔다. 우리는 이전에이를 apoptogenic 요인 12,14의 릴리스에 의한 apoptosis를 유도 암세포에 선택적으로 mitochondria를 불안정하게하는 자연 PST를보고했습니다. 그것은 가능성이 높습니다 그 같은 메커니즘을 통해 JCTH-4 막으로, JCTH-4 (TMRM 염색법 (그림 5A)와 볼, 그리고 SH-SY5Y 세포 격리 mitochondria의 ROS의 생성을 증가...

<table border="1"><tbody><tr><td> 자재 명칭 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> SH-SY5Y 세포주 </td><td> ATCC </td><td> CRL-2266 </td></tr><tr><td> Dulbecco의 수정일 이글스 중 F-12 햄 </td><td> 시그마 - 올드 리치 </td><td> 51448C </td></tr><tr><td> 태아 소 혈청 </td><td> Gibco BRL </td><td> 16000-044 </td></tr><tr><td> MCF7 세포주 </td><td> ATCC </td><td> HTB-22 </td></tr><tr><td> RPMI-1640 배지 </td><td> 시그마 - 올드 리치 </td><td> R 0883 </td></tr><tr><td> 분명히 정상적인 인간 태아의 fibroblast 세포주 (NFF) </td><td> 의학 연구를위한 Coriell 연구소 </td><td> AG04431B </td></tr><tr><td> Dulbecco의 수정일 이글의 중간, 높음 포도당 매체 </td><td> 써모 과학 </td><td> SH30022.01 </td></tr><tr><td> Tamoxifen의 구연산 염 </td><td> 시그마 - 올드 리치 </td><td> T9262 </td&gt; </tr><tr><td> 35mm 유리 바닥 문화 요리 </td><td> MatTek </td><td> P35G-014-C </td></tr><tr><td> Leica DMI6000 B 거꾸로 현미경 </td><td> Leica 마이크로 시스템즈 </td><td> N / A </td></tr><tr><td> Hoechst 33342 염색 </td><td> 분자 프로브 </td><td> H3570 </td></tr><tr><td> Leica DM의 IRB는 형광 현미경을 거꾸로 </td><td> Leica 마이크로 시스템즈 </td><td> N / A </td></tr><tr><td> Annexin V AlexaFluor-488 </td><td> Invitrogen </td><td> A13201 </td></tr><tr><td> Trypan 블루 용액 </td><td> 시그마 - 올드 리치 </td><td> T8154-20ML </td></tr><tr><td> 혈구계 </td><td> 피셔 과학 </td><td> 267,110 </td></tr><tr><td> 서부 표준시-1 시약 </td><td> 로슈 응용 과학 </td><td> 11644807001 </td></tr><tr><td> Wallac 빅터 3 1420 Multilabel 카운터 </td><td> PerkinElmer </td><td> 1420-011 </td></tr><tr><td> Tetramethylrhodamine 메틸 에스테르 (TMRM) </td><td> Gibco BRL </td><td> T-668 </td></tr><tr><td> 유리 조직 연삭기 </td><td> 피셔 과학 </td><td> K8885300-0002 </td></tr><tr><td> BioRad 단백질 분석 </td><td> 바이오 방사선 연구소 </td><td> 양식 500 0001Bottom </td></tr><tr><td> Amplex 레드 </td><td> Invitrogen </td><td> A12222 </td></tr><tr><td> 양고추냉이 퍼옥시데이즈 (HRP) </td><td> 시그마 - 올드 리치 </td><td> P8125 </td></tr><tr><td> SpectraMax 쌍둥이 자리 XPS </td><td> 분자 장치 </td><td> 3,126,666 </td></tr><tr><td> 안티 LC3 항체는 토끼의 사육 </td><td> Novus 체액 </td><td> NB100-2220 </td></tr><tr><td> 안티 마우스 HRP-복합 이차 항체 </td><td> Abcam </td><td> ab6728 </td></tr><tr><td> 안티 - 토끼 HRP-복합 이차 항체 </td><td> Abcam </td><td> ab6802 </td></tr><tr><td> Chemiluminescence 퍼옥시데이즈 기판 </td><td> 시그마 - 올드 리치 </td><td> CPS160 </td></tr><tr><td> Monodansylcadaverine </td><td> 시그마 - 올드 리치 </td><td> 30,432 </td></tr></tbody></table>

enhancement of apoptotic and autophagic induction by a novel synthetic c-1 analogue of 7-deoxypancratistatin in human breast adenocarcinoma and neuroblastoma cells with tamoxifen

유방암은 북미에서 여인 중에 가장 흔한 암 중 하나입니다. 이온화 방사선 등 많은 현재의 안티 - 암 치료, DNA 손상을 통해 apoptosis를 유도. 불행히도, 이러한 치료는 암세포가 아닌 선택이며, 정상 세포에서 유사한 독성을 생산하고 있습니다. 우리는 천연 복합 pancratistatin (PST)에 의해 암 세포에서 apoptosis의 선택적 유도를보고했습니다. 최근 소설 태평양 표준시 아날로그, 7 deoxypancratistatin (JCTH-4)의 C-아세 톡시 메틸 유도체는 드 노보 합성에 의해 생산과 몇 가지 암 세포 라인에 비교 선택적 apoptosis를 유도하는 활동을 전시했다. 최근 autophagy는 화학 요법에 대한 반응으로 프로 생존과 프로 사망 메커니즘 양쪽으로 malignancies에 연루되었습니다. Tamoxifen (TAM)는 결국에 수많은 암의 프로 생존 autophagy의 유도를 보여주었다. 본 연구에서는 혼자와 TAM와 함께 JCTH-4의 효능은 인간의 유방 cance에서 세포 죽음을 유도하는R (MCF7)과 neuroblastoma (SH-SY5Y) 전지 평가되었다. 혼자 JCTH-4 autophagy의 일부 유도와 apoptosis의 상당 유도를 일으킨 반면 혼자 TAM은 autophagy 유도하지만, 볼품없는 세포의 죽음. 흥미롭게도, 조합 치료 apoptotic과 autophagic 유도의 급격한 증가를 굴복. 우리는 촬영된 현미경을 사용하여 조합 치료 TAM 유발 autophagy를 겪고 MCF7 세포에서는 시간 의존 형태학의 변화, JCTH-4-유도된 apoptosis와 autophagy 및 가속 세포 죽음을 모니터링. 우리는이 암 세포의 mitochondrial 타겟팅을 사용하여 apoptosis / autophagy를 유도하기 위해 이러한 화합물을 증명하고있다. 중요한 것은 이러한 치료 noncancerous 인간 섬유아 세포의 생존에 영향을주지 않았다. 따라서 이러한 결과는 JCTH-4 TAM과 결합된이 유방암과 neuroblastoma 세포에 대한 안전하고 매우 강력한 항정 암 치료로 사용될 수 있음을 나타냅니다.

Watch this Scientific Journal Video about Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma 

Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma Cells with Tamoxifen

우리는 네이티브 pancratistatin 같은 비교 안티 - 암 활동과 pancratistatin의 소설 아날로그를 합성했습니다, 흥미롭게도, tamoxifen과 조합 치료 noncancerous 섬유아 세포에 대한 최소한의 효과와 대상 mitochondrial에 의한 apoptotic 및 autophagic 유도의 급격한 향상이 나왔고. 따라서, tamoxifen와 함께 JCTH-4는 안전한 안티 암 치료를 제공할 수 있습니다.

인간 유방 선암과 Tamoxifen과 Neuroblastoma 셀의 7-deoxypancratistatin의 소설 합성 C-1 아날로그에 의한 Apoptotic 및 Autophagic 유도의 향상

Breast cancer is one of the most common cancers amongst women in North America. Many current anti-cancer treatments, including ionizing radiation, induce ...

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18.6K 조회수.  University of Windsor. 우리는 네이티브 pancratistatin 같은 비교 안티 - 암 활동과 pancratistatin의 소설 아날로그를 합성했습니다, 흥미롭게도, tamoxifen과 조합 치료 noncancerous 섬유아 세포에 대한 최소한의 효과와 대상 mitochondrial에 의한 apoptotic 및 autophagic 유도의 급격한 향상이 나왔고. 따라서, tamoxifen와 함께 JCTH-4는 안전한 안티 암 치료를 제공할 수 있습니다.

비디오: Enhancement of Apoptotic and Autophagic Induction by a Novel Synthetic C-1 Analogue of 7-deoxypancratistatin in Human Breast Adenocarcinoma and Neuroblastoma Cells with Tamoxifen

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human pulp-derived stem cells (hDPSCs) possess high proliferation potential with multi-lineage differentiation capacity compare to the ordinary source of adult stem cells1-8; therefore, hDPSCs could be the good candidates for autologous transplantation in tissue engineering and regenerative medicine. Along with these benefits, possessing the mesenchymal stem cells (MSC) features, such as immunolodulatory effect, make hDPSCs more valuable, even in the case of allograft transplantation6,9,10. Therefore, the primary step for using this source of stem cells is to select the best protocol for isolating hDPSCs from pulp tissue. In order to achieve this goal, it is crucial to investigate the effect of various isolation conditions on different cellular behaviors, such as their common surface markers &amp; also their differentiation capacity.
Thus, here we separate human pulp tissue from impacted third molar teeth, and then used both existing protocols based on literature, for isolating hDPSCs,11-13 i.e. enzymatic dissociation of pulp tissue (DPSC-ED) or outgrowth from tissue explants (DPSC-OG). In this regards, we tried to facilitate the isolation methods by using dental diamond disk. Then, these cells characterized in terms of stromal-associated Markers (CD73, CD90, CD105 &amp; CD44), hematopoietic/endothelial Markers (CD34, CD45 &amp; CD11b), perivascular marker, like CD146 and also STRO-1. Afterwards, these two protocols were compared based on the differentiation potency into odontoblasts by both quantitative polymerase chain reaction (QPCR) &amp; Alizarin Red Staining. QPCR were used for the assessment of the expression of the mineralization-related genes (alkaline phosphatase; ALP, matrix extracellular phosphoglycoprotein; MEPE &amp; dentin sialophosphoprotein; DSPP).14

The method described isolation and characterization of human Dental Pulp Stem Cells (hDPSCs) by using either enzymatic dissociation of pulp (DPSC-ED) or direct outgrowth of stem cells from pulp tissue explants (DPSC-OG). Then followed by in vitro comparative differentiation of both types of hDPSCs into odontoblasts.

인간 치과 펄프의 줄기 세포의 분리, 특성화 및 비교 차별화 두 가지 방법을 사용하여 영구 치아에서 파생

Developing wisdom teeth are easy-accessible source of stem cells during the adulthood which could be obtained by routine orthodontic treatments. Human ...

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Cell Death Associated with Abnormal Mitosis Observed by Confocal Imaging in Live Cancer Cells

Phenanthrene derivatives acting as potent PARP1 inhibitors prevented the bi-focal clustering of supernumerary centrosomes in multi-centrosomal human cancer cells in mitosis. The phenanthridine PJ-34 was the most potent molecule. Declustering of extra-centrosomes causes mitotic failure and cell death in multi-centrosomal cells. Most solid human cancers have high occurrence of extra-centrosomes. The activity of PJ-34 was documented in real-time by confocal imaging of live human breast cancer MDA-MB-231 cells transfected with vectors encoding for fluorescent &gamma;-tubulin, which is highly abundant in the centrosomes and for fluorescent histone H2b present in the chromosomes. Aberrant chromosomes arrangements and de-clustered &gamma;-tubulin foci representing declustered centrosomes were detected in the transfected MDA-MB-231 cells after treatment with PJ-34. Un-clustered extra-centrosomes in the two spindle poles preceded their cell death. These results linked for the first time the recently detected exclusive cytotoxic activity of PJ-34 in human cancer cells with extra-centrosomes de-clustering in mitosis, and mitotic failure leading to cell death. According to previous findings observed by confocal imaging of fixed cells, PJ-34 exclusively eradicated cancer cells with multi-centrosomes without impairing normal cells undergoing mitosis with two centrosomes and bi-focal spindles. This cytotoxic activity of PJ-34 was not shared by other potent PARP1 inhibitors, and was observed in PARP1 deficient MEF harboring extracentrosomes, suggesting its independency of PARP1 inhibition. Live confocal imaging offered a useful tool for identifying new molecules eradicating cells during mitosis.

The cytotoxic activity of the phenanthridine PJ-34 in cancer cells undergoing mitosis was documented in real time by live confocal imaging. PJ-34 eradicated human breast cancer MDA-MB-231 cells harboring extra-centrosomes in mitosis. Unlike normal bi-focal mitosis, the extra-centrosomes were not clustered in the two spindle poles in the presence of PJ-34.

비정상적인 유사 분열과 관련된 세포의 죽음은 라이브 암 세포의 공 촛점 이미징에 의해 관찰

Phenanthrene derivatives acting as potent PARP1 inhibitors prevented the bi-focal clustering of supernumerary centrosomes in multi-centrosomal human ...

Initiation of Metastatic Breast Carcinoma by Targeting of the Ductal Epithelium with Adenovirus-Cre: A Novel Transgenic Mouse Model of Breast Cancer

Breast cancer is a heterogeneous disease involving complex cellular interactions between the developing tumor and immune system, eventually resulting in exponential tumor growth and metastasis to distal tissues and the collapse of anti-tumor immunity. Many useful animal models exist to study breast cancer, but none completely recapitulate the disease progression that occurs in humans. In order to gain a better understanding of the cellular interactions that result in the formation of latent metastasis and decreased survival, we have generated an inducible transgenic mouse model of YFP-expressing ductal carcinoma that develops after sexual maturity in immune-competent mice and is driven by consistent, endocrine-independent oncogene expression. Activation of YFP, ablation of p53, and expression of an oncogenic form of K-ras was achieved by the delivery of an adenovirus expressing Cre-recombinase into the mammary duct of sexually mature, virgin female mice. Tumors begin to appear 6 weeks after the initiation of oncogenic events. After tumors become apparent, they progress slowly for approximately two weeks before they begin to grow exponentially. After 7-8 weeks post-adenovirus injection, vasculature is observed connecting the tumor mass to distal lymph nodes, with eventual lymphovascular invasion of YFP+ tumor cells to the distal axillary lymph nodes. Infiltrating leukocyte populations are similar to those found in human breast carcinomas, including the presence of &#945;&#946; and &#947;&#948; T cells, macrophages and MDSCs. This unique model will facilitate the study of cellular and immunological mechanisms involved in latent metastasis and dormancy in addition to being useful for designing novel immunotherapeutic interventions to treat invasive breast cancer.

Activation of latent mutations with adenovirus-Cre into the mammary ductal system results in a clinically relevant metastatic breast cancer. Incorporation of a YFP promoter allows tracking of distal metastatic tumor cells. This model is useful to study latent metastasis, anti-tumor immunity, and for designing novel immunotherapies to treat breast cancer.

유방암의 소설 형질 전환 마우스 모델 : 아데노 바이러스 크레와 유관 상피의 대상으로 전이성 유방암의 개시

Breast cancer is a heterogeneous disease involving complex cellular interactions between the developing tumor and immune system, eventually resulting in ...

Profiling of Estrogen-regulated MicroRNAs in Breast Cancer Cells

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen receptors (ERs), ER&#945;, and ER&#946;. ER&#945; is frequently upregulated in breast cancer and drives the proliferation of breast cancer cells. The ERs function as transcription factors and regulate gene expression. Whereas ER&#945;&#39;s regulation of protein-coding genes is well established, its regulation of noncoding microRNA (miRNA) is less explored. miRNAs play a major role in the post-transcriptional regulation of genes, inhibiting their translation or degrading their mRNA. miRNAs can function as oncogenes or tumor suppressors&#160;and are also promising biomarkers. Among the miRNA assays available, microarray and quantitative real-time polymerase chain reaction (qPCR) have been extensively used to detect and quantify miRNA levels. To identify miRNAs regulated by estrogen signaling in breast cancer, their expression in ER&#945;-positive breast cancer cell lines were compared before and after estrogen-activation using both the &#181;Paraflo-microfluidic microarrays and Dual Labeled Probes-low density arrays. Results were validated using specific qPCR assays, applying both Cyanine dye-based and Dual Labeled Probes-based chemistry. Furthermore, a time-point assay was used to identify regulations over time. Advantages of the miRNA assay approach used in this study is that it enables a fast screening of mature miRNA regulations in numerous samples, even with limited sample amounts. The layout, including the specific conditions for cell culture and estrogen treatment, biological and technical replicates, and large-scale screening followed by in-depth confirmations using separate techniques, ensures a robust detection of miRNA regulations,&#160;and eliminates false positives and other artifacts. However, mutated or unknown miRNAs, or regulations at the primary and precursor transcript level, will not be detected. The method presented here represents a thorough investigation of estrogen-mediated miRNA regulation.

Molecular signaling through both estrogen and microRNAs are critical in breast cancer development and growth. Estrogen activates the estrogen receptors, which are transcription factors. Many transcription factors can regulate the expression of microRNAs, and estrogen-regulated microRNAs can be profiled using different large-scale techniques.

유방암 세포에서 에스트로겐 조절 마이크로 RNA의 프로파일

Estrogen plays vital roles in mammary gland development and breast cancer progression. It mediates its function by binding to and activating the estrogen ...

Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells

Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation assay (hereafter called soft agar assay), which measures the ability of cells to proliferate in semi-solid matrices, remain a hallmark of cancer research. A key advantage of this technique over conventional 2D monolayer or 3D spheroid cell culture assays is the close mimicry of the 3D cellular environment to that seen in vivo. Importantly, the soft agar assay also provides an ideal tool to rigorously test the effects of novel compounds or treatment conditions on cell proliferation and migration. Additionally, this assay enables the quantitative assessment of cell transformation potential within the context of genetic perturbations. We recently identified peptidylarginine deiminase 2 (PADI2) as a potential breast cancer biomarker and therapeutic target. Here we highlight the utility of the soft agar assay for preclinical anti-cancer studies by testing the effects of the PADI inhibitor, BB-Cl-amidine (BB-CLA), on the tumorigenicity of human ductal carcinoma in situ (MCF10DCIS) cells.

Here, we document the use of the soft agar colony formation assay to test the effects of a peptidylarginine deiminase (PADI) enzyme inhibitor, BB-Cl-amidine, on breast cancer tumorigenicity in vitro.

소프트 한천 콜로니 형성 분석의 활용은 유방암 세포에서 종양 유발의 억제를 확인하는

Given the inherent difficulties in investigating the mechanisms of tumor progression in vivo, cell-based assays such as the soft agar colony formation ...

Systems Biology of Metabolic Regulation by Estrogen Receptor Signaling in Breast Cancer

With the advent of the -omics approaches our understanding of the chronic diseases like cancer and metabolic syndrome has improved. However, effective mining of the information in the large-scale datasets that are obtained from gene expression microarrays, deep sequencing experiments or metabolic profiling is essential to uncover and then effectively target the critical regulators of diseased cell phenotypes. Estrogen Receptor &#945; (ER&#945;) is one of the master transcription factors regulating the gene programs that are important for estrogen responsive breast cancers. In order to understand to role of ER&#945; signaling in breast cancer metabolism we utilized transcriptomic, cistromic and metabolomic data from MCF-7 cells treated with estradiol. In this report we described generation of samples for RNA-Seq, ChIP-Seq and metabolomics experiments and the integrative computational analysis of the obtained data. This approach is useful in delineating novel molecular mechanisms and gene regulatory circuits that are regulated by a particular transcription factor which impacts metabolism of normal or diseased cells.

Integration of data from genome-wide sequencing experiments and metabolomics experiments is a challenge. In this paper we report, for the first time, generation, analysis and integration of transcriptome, cistrome and metabolome data from breast cancer cells treated with estradiol.

유방암에서 에스트로겐 수용체 신호 전달에 의해 대사 규제 시스템 생물학

With the advent of the -omics approaches our understanding of the chronic diseases like cancer and metabolic syndrome has improved. However, effective ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells

Mesenchymal Stromal Cells (MSCs) are tissue homeostatic and immune modulatory cells that have shown beneficial effects in kidney diseases and transplantation. Perivascular Stromal Cells (PSCs) share characteristics with bone marrow MSCs (bmMSCs). However, they also possess, most likely due to local imprinting, tissue-specific properties and play a role in local tissue homeostasis. This tissue specificity may result in tissue specific repair, also within the human kidney. We previously showed that human kidney PSCs (kPSCs) have enhanced kidney epithelial wound healing whereas bmMSCs did not have this potential. Moreover, kPSCs can ameliorate kidney injury in vivo. Therefore, kPSCs constitute an interesting source for cell therapy, particularly for kidney diseases and renal transplantation. Here we show the detailed isolation and culture method for kPSCs from transplant-grade human kidneys based on whole-organ perfusion of digestive enzymes via the renal artery and enrichment for the perivascular marker NG2. In this way, large cell quantities can be obtained that are suitable for cellular therapy.

Here we present a novel clinical grade isolation and culture method for kidney Perivascular Stromal Cells (kPSCs) based on whole organ perfusion with digestive enzymes and NG2-cell enrichment. With this method, it is possible to acquire sufficient cell numbers for cellular therapy.

인간의 신장 혈관 주위 Stromal 세포에 대 한 새로운 임상 등급 격리 방법

Mesenchymal Stromal Cells (MSCs) are tissue homeostatic and immune modulatory cells that have shown beneficial effects in kidney diseases and ...

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a wide variety of disorders including metabolic diseases, premature aging syndromes, and neurodegenerative diseases such as Alzheimer's disease (AD) and Parkinson's disease (PD). Maintenance of mitochondrial health depends on mitochondrial biogenesis and the efficient clearance of dysfunctional mitochondria through mitophagy. Experimental methods to accurately detect autophagy/mitophagy, especially in animal models, have been challenging to develop. Recent progress towards the understanding of the molecular mechanisms of mitophagy has enabled the development of novel mitophagy detection techniques. Here, we introduce several versatile techniques to monitor mitophagy in human cells, Caenorhabditis elegans (e.g., Rosella and DCT-1/ LGG-1 strains), and mice (mt-Keima). A combination of these mitophagy detection techniques, including cross-species evaluation, will improve the accuracy of mitophagy measurements and lead to a better understanding of the role of mitophagy in health and disease.

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

Mitophagy, the process of clearing damaged mitochondria, is necessary for mitochondrial homeostasis and health maintenance. This article presents some of the latest mitophagy detection methods in human cells, Caenorhabditis elegans, and mice.

생체 외에서 그리고 Vivo에서 인간 세포, C. 선 충, 쥐에서 Mitophagy의 검출

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a ...

Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry

Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the development of obesity-associated complications such as type 2 diabetes. To quantitatively and qualitatively investigate the immune cell subsets in human AT deposits, we have developed a flow cytometry approach. The stromal vascular fraction (SVF), containing the immune cells, is isolated from subcutaneous and visceral AT biopsies by collagenase digestion. Adipocytes are removed after centrifugation. The SVF cells are stained for multiple membrane-bound markers selected to differentiate between immune cell subsets and analyzed using flow cytometry. As a result of this approach, pro- and anti-inflammatory macrophage subsets, dendritic cells (DCs), B-cells, CD4+ and CD8+ T-cells, and NK cells can be detected and quantified. This method gives detailed information about immune cells in AT and the amount of each specific subset. Since there are numerous fluorescent antibodies available, our flow cytometry approach can be adjusted to measure various other cellular and intracellular markers of interest.

This article describes a method to analyze immune cell content of adipose tissue by isolation of immune cells from adipose tissue and subsequent analysis using flow cytometry.