Name: methods for analyzing the impacts of natural uranium on in vitro osteoclastogenesis
Uploaded: 2018-01-30T13:00:00.000+00:00
Duration: 9 min 51 s
Description: Watch this Scientific Journal Video about Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis at JoVE.com

저자 Chantal Cros 도움이 기술 지원에 대해 감사 하 고 싶습니다. 이 연구는에서 교부 금에 의해 투자 되었다는 "Commissariat à l'Energie Atomique 외 보조 에너지 대안" (URANOs-프로그램 횡단 de Toxicologie du CEA와 CPRR CEA-AREVA), ANR에서 (우라늄의 독성: biomineralization의 다단계 접근 뼈, ANR-16-CE34-0003에서에서 처리). 이 작품 또한 좋은 소피아 앙 티 폴리스는 CNRS의 대학에 의해 지원 되었다.

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저자 공개할 게 없다

우리가 알기로 이것이 처음으로 뼈 세포를 resorbing에 자연 우라늄의 효과 연구를 목표로 자세한 절차 설명 하. 이 방법은 뼈 생리학에 우라늄 영향의 더 나은 이해를 달성 하는 데 유용 되며 우라늄 chelators의 심사에 대 한 흥미로운 새로운 도구를 제공할 수 있습니다. 또한, 우리는 여기에 설명 된 프로토콜 osteoclatogenesis에 다른 중 금속의 영향을 연구에 적용 될 수 믿습니다.
그...

우라늄은 자연 발생 방사능 요소 토양, 공기 및 물;에 따라서, 동물과 인간은 그들의 규정식에 있는 자연 우라늄에 노출 됩니다. 자연 소스 뿐만 아니라 우라늄 어떤 환경에서 그것의 풍부를 증가 anthropogenic 활동에서 유래. 우라늄 화학 및 방사능 위험 포즈. 그러나, (이 동위 원소 혼합물 포함 99.27 238U, 0.72 235U, 그리고 0.006 234U) 자연 우라늄 낮은 특정 활동 (25.103 Bq.g-1)가 있기 때문에 건강에 미치는 영향에 기인의 화학 독성입니다.
무엇이 든 해당 항목 경로 (흡입, 섭취, 또는 피부 노출), 대부분 우라늄의 배설물 제거는 시체를 입력 하 고 단지 작은 부분에 도달 하면 조직의 순환. 혈액에서 우라늄의 약 67%에 신장에 의해 필터링 되 고 24 h1내 소변에서 시체를 나뭇잎. 나머지는 주로 신장과 뼈, 우라늄의 독성2,,34의 두 가지 주요 표적 기관에 입금 됩니다. 여러 연구 탐험 실시 되어 골격 우라늄 장기 보존2,,34,,56의 주 사이트 발견 되었습니다, 때문에 골 생리학7에 우라늄의 효과.
뼈는 일생에 걸쳐 지속적으로 리 모델링 광물 화 된 조직 이다. 뼈는 복잡 한 프로세스 전문된 셀 형식에 따라 달라 집니다 이며 대부분의 두 단계로 구성 됩니다: osteoclasts 하 여 기존의 오래 된 매트릭스의 재흡수 osteoblasts 드 노 보 뼈 구조에 의해 옵니다. Osteoclasts 있습니다 큰 多 어디 그들은8뼈를 연결 하는 재흡수 사이트로 마이그레이션하는 조 혈 근원의 전조 세포의 융해에서 유래. 그들의 첨부 파일은 그들의 골격9의 광범위 한 개편으로 동시에 발생합니다. 이 개편은 셀과로 osteoclast secretes 양성자, hydroxyapatite, 및의 저하에 관련 된 프로 테아의 해체로 이어지는 뼈 표면 사이 격리 된 구획의 설립을 위한 필요는 유기 매트릭스입니다. 결과 저하 제품 endocytosed, 뼈 표면에 반대 막 영역에 셀을 통해 수송 되며, 분 비, transcytosis10,11을 불리는 과정.
Vivo에서 그리고 생체 외에서 학문에서 결과 우라늄 뼈 형성 억제와 osteoblasts7,12의 활동을 변경 나타냅니다. 대조적으로, 뼈 재흡수 및 osteoclasts에 우라늄의 효과 제대로 탐험 되었습니다 있다. 여러 vivo에서 연구 쥐 또는 쥐13,14uranyl 질 산의 관리 후 뼈 재흡수의 향상을 보고 있다. 또한, 역학 조사는 식 수를 통해 우라늄 섭취의 증가 경향이 남자15뼈 재흡수 표시자의 혈 청 수준에 있는 증가와 관련 된 제안 했다. 함께 찍은, 이러한 결과 우라늄, 뼈에 축적 되어 뼈 재흡수를 홍보할 수 있는 결론으로 이끌어 냈다. 그러나, 우라늄의이 잠재적인 효과에 관련 된 세포 메커니즘 미결 문제 남아 있다. 이러한 이유로, 우리는 뼈 세포 resorbing의 동작에 우라늄의 영향을 검사 하기로 결정 했다.
여기, 우리는 우리가 설정한 프로토콜 설명 하 특성화 고와 osteoclasts 차별화 및 resorptive 활동 사전 osteoclasts 생존에 자연 우라늄의 효과 계량. 여기에 설명 된 실험 RAW 264.7 murine 변형 된 macrophage 셀 라인,는 쉽게 osteoclasts 때 4 ~ 5 일에 대 한 cytokine RANKL의 존재 양식으로 분화 할 수 있다 하 고 고전적인 연구에 사용 되는 할 osteoclast 차별화 및 기능16. 개발 절차는 신뢰할 수 있는, 높은 재현성 결과는 완전히 기본 osteoclasts에 적용. 모든 이러한 이유로, 우리는이 방법론은 뼈에 우라늄 독성에 관련 된 분자 메커니즘을 더 잘 이해 해 줘 유용 믿습니다. 또한, 우리는이 이렇게 새로운 우라늄 킬레이트 화 대리인 확인을 위한 검사 도구로 적응 될 수 있다고 생각 합니다.

<table><tbody><tr><td>DMEM</td><td>Lonza</td><td>BE12-604F</td><td></td></tr><tr><td>&amp;alpha;-MEM</td><td>Lonza</td><td>BE12-169F</td><td></td></tr><tr><td>EMEM without phenol red</td><td>Lonza</td><td>12-668E</td><td></td></tr><tr><td>Water for cell culture</td><td>Lonza</td><td>BE17-724F</td><td></td></tr><tr><td>PBS</td><td>Sigma-Aldrich</td><td>D8537</td><td></td></tr><tr><td>Penicillin-Streptomycin solution</td><td>Sigma-Aldrich</td><td>P4333</td><td></td></tr><tr><td>&amp;nbsp;L-Glutamine solution</td><td>Sigma-Aldrich</td><td>G7513</td><td></td></tr><tr><td>Trypan Blue Solution 0.4%</td><td>Sigma-Aldrich</td><td>T8154</td><td></td></tr><tr><td>HyClone fetal bovine serum</td><td>GE Life Sciences</td><td>SH30071.03</td><td></td></tr><tr><td>7.5% sodium bicarbonate aqueous solution</td><td>Sigma-Aldrich</td><td>S8761</td><td></td></tr><tr><td>Acid Phosphatase, Lekocyte (TRAP) kit</td><td>Sigma-Aldrich</td><td>387A</td><td></td></tr><tr><td>Thiazolyl Blue Tetrazolium Bromide (MTT) powder</td><td>Sigma-Aldrich</td><td>M5655</td><td></td></tr><tr><td>Dimethyl sulfoxide</td><td>Sigma-Aldrich</td><td>D5879</td><td></td></tr><tr><td>Alizarin Red S sodium salt, 1% w/v aq. sol.</td><td>Alfa Aeros</td><td>42746</td><td></td></tr><tr><td>Osteoassay bone resorption plates, 24 well plates</td><td>Corning Life Sciences</td><td>3987</td><td></td></tr><tr><td>Multiwell 24 well plates</td><td>Falcon</td><td>353504</td><td></td></tr><tr><td>Flask 75 cm&lt;sup&gt;2&lt;/sup&gt;</td><td>Falcon</td><td>353133</td><td></td></tr><tr><td>Polypropylene Conical Tubes 50 ml</td><td>Falcon</td><td>352070</td><td></td></tr><tr><td>Cell scrapers 30 cm</td><td>TPP</td><td>90003</td><td></td></tr></tbody></table>

methods for analyzing the impacts of natural uranium on in vitro osteoclastogenesis

우라늄은 뼈 생리학 방해을 보여줘 왔다 그리고 그것은 잘이 금속 뼈에 축적 되어 설립. 그러나, 작은 뼈 세포의 행동에 자연 우라늄의 효과 대 한 알려져 있다. 특히, osteoclasts 뼈 매트릭스의 재흡수에 대 한 책임 셀에 우라늄의 충격은 문서화 되어 있지. 이 문제를 조사, 우리 osteoclast 선구자의 모델로 자연 우라늄의 원본과 murine RAW 264.7 세포 라인으로 uranyl 아세테이트를 사용 하 여 새 프로토콜을 설립 했다. 여기, 우리가 상세한 osteoclast 선구자에 우라늄 세포 독성을 테스트 하 고는 osteoclastogenesis에 그리고 성숙한 osteoclasts의 resorbing 기능에 미치는 영향을 평가 하는 데 필요한 모든 분석 실험. 우리가 개발한 조건, 특히 uranyl 포함 된 문화 미디어의 준비와 RAW 264.7 시드 셀 안정적이 고 높은 생식 결과를 얻을 수 있습니다. 또한, 우리 osteoclasts의 크기 또는 resorbed 매트릭스의 비율 등 다양 한 매개 변수 분석을 촉진 하기 위하여 소프트웨어 도구의 사용을 최적화 했습니다.

주석산 저항 산 성 인산 가수분해 효소 얼룩 3 개 이상의 핵 데 큰 보라색 셀 osteoclasts 시각화 사용 되었다. RANKL의 경작 된 RAW 264.7 세포에서 얻은 osteoclasts의 대표 이미지 그리고 uranyl 이온 그림 1에 나와 있습니다. Osteoclasts 우라늄에 대 한 응답에서의 수와 크기에 변화 전체 우물의 합성 이미지와 확대 사진을 쉽게 볼 수 있습니다.
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Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

우라늄은 골 대사에 영향을 미칠으로 알려져 있다. 여기, 선물이 생존, 분화, 및 osteoclasts 뼈 재흡수를 담당 하 고 세포의 기능에 자연 우라늄 노출의 효과 조사 하기 위한 프로토콜.

생체 외에서 Osteoclastogenesis에 자연 우라늄의 영향을 분석 하기 위한 방법

Uranium has been shown to interfere with bone physiology and it is well established that this metal accumulates in bone. However, little is known about ...

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JoVE Journal

Developmental Biology

Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

8.1K 조회수.  Université Nice Sophia-Antipolis. 우라늄은 골 대사에 영향을 미칠으로 알려져 있다. 여기, 선물이 생존, 분화, 및 osteoclasts 뼈 재흡수를 담당 하 고 세포의 기능에 자연 우라늄 노출의 효과 조사 하기 위한 프로토콜.

비디오: Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted through ion exchange. The resultant production bleed water (PBW) contains contaminants such as arsenic and other heavy metals. Samples of PBW from an active ISR uranium facility were treated with cupric oxide nanoparticles (CuO-NPs). CuO-NP treatment of PBW reduced priority contaminants, including arsenic, selenium, uranium, and vanadium. Untreated and CuO-NP treated PBW was used as the liquid component of the cell growth media and changes in viability were determined by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay in human embryonic kidney (HEK 293) and human hepatocellular carcinoma (Hep G2) cells. CuO-NP treatment was associated with improved HEK and HEP cell viability. Limitations of this method include dilution of the PBW by growth media components and during osmolality adjustment as well as necessary pH adjustment. This method is limited in its wider context due to dilution effects and changes in the pH of the PBW which is traditionally slightly acidic&#160;however; this method could have a broader use assessing CuO-NP treatment in more neutral waters.

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

우라늄에서 구리 산화물 나노 입자에 의한 미량 원소의 제거<em&gt; 현장에서</em&gt; 복구 블리드 물과 세포 생존에 미치는 영향

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

연구

환경과학

Development of a Human Preclinical Model of Osteoclastogenesis from Peripheral Blood Monocytes Co-cultured with Breast Cancer Cell Lines

The crosstalk between tumor cells and bone cells in the bone microenvironment is crucial to understanding the mechanism of bone metastasis formation. We developed an in vitro fully human preclinical model of a co-culture of breast cancer cells and monocytes undergoing differentiation towards osteoclasts. We optimized a model of osteoclastogenesis starting from a sample of peripheral blood collected from healthy donors. Peripheral blood mononuclear cells (PBMCs) were first separated by density gradient centrifugation, seeded at a high density and induced to differentiate by adding two growth factors (GFs): receptor activator of nuclear factor-&#954;B ligand (RANKL) and macrophage colony-stimulating factor (MCSF). The cells were left in culture for 14 days and then fixed and analyzed by downstream analysis. In osteolytic bone metastases, one of the effects of cancer cell arrival in bone is the induction of osteoclastogenesis. We thus challenged our model with co-cultures of breast cancer cells to study the differentiation power of cancer cells with respect to GFs. A straightforward way of studying cancer cell-osteoclast interaction is to perform indirect co-cultures based on the use of conditioned medium collected from breast cancer cell cultures and mixed with fresh medium. This mixture is then used to induce osteoclast differentiation. We also optimized a method of direct co-culture in which cancer cells and monocytes undergoing differentiation share the medium and exchange secreted factors. This is a significant improvement over the original indirect co-culture method as researchers can observe the reciprocal interactions of the two cell types and perform downstream analyses for both cancer cells and osteoclasts. This method enables us to study the effect of drugs on the metastatic bone microenvironment and to seed cell lines other than those derived from breast cancer. The model can also be used to study other diseases such as osteoporosis or other bone conditions.

This protocol describes development of an in vitro human preclinical model of osteoclastogenesis from peripheral blood monocytes cultured with breast cancer cell lines to mimic the cancer cell-osteoclast interaction. The model could be used to further our understanding of bone metastasis formation and improve therapeutic options.

말 초 혈액 Monocytes 유 방 암 세포 라인 공동 경작에서 Osteoclastogenesis의 인간의 전 임상 모델의 개발

The crosstalk between tumor cells and bone cells in the bone microenvironment is crucial to understanding the mechanism of bone metastasis formation. We ...

암 연구학

A RANKL-based Osteoclast Culture Assay of Mouse Bone Marrow to Investigate the Role of mTORC1 in Osteoclast Formation

Osteoclasts are unique bone-resorbing cells that differentiate from the monocyte/macrophage lineage of bone marrow. Dysfunction of osteoclasts may result in a series of bone metabolic diseases, including osteoporosis. To develop pharmaceutical targets for the prevention of pathological bone mass loss, the mechanisms by which osteoclasts differentiate from precursors must be understood. The ability to isolate and culture a large number of osteoclasts in vitro is critical in order to determine the role of specific genes in osteoclast differentiation. Inactivation of the mammalian/mechanistic target of rapamycin complex 1 (TORC1) in osteoclasts can decrease osteoclast number and increase bone mass; however, the underlying mechanisms require further study. In the present study, a RANKL-based protocol to isolate and culture osteoclasts from mouse bone marrow and to study the influence of mTORC1 inactivation on osteoclast formation is described. This protocol successfully resulted in a large number of giant osteoclasts, typically within one week. Deletion of Raptor impaired osteoclast formation and decreased the activity of secretory tartrate-resistant acid phosphatase, indicating that mTORC1 is critical for osteoclast formation.

This manuscript describes a protocol to isolate and culture osteoclasts in vitro from mouse bone marrow, and to study the role of the mammalian/mechanistic target of rapamycin complex 1 in osteoclast formation.

RANKL 기반 Osteoclast 문화 분석 결과의 마우스 골 수 mTORC1 Osteoclast 형성에의 역할을 조사 하

Osteoclasts are unique bone-resorbing cells that differentiate from the monocyte/macrophage lineage of bone marrow. Dysfunction of osteoclasts may result ...

발생학

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

소설<em&gt; 생체</em&gt; 유전자 전달 기술과<em&gt; 시험관</em근골격계 질환의 뼈 손실의 연구를위한&gt; 셀 기반 분석 실험

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

의학

Effect of Anti-c-fms Antibody on Osteoclast Formation and Proliferation of Osteoclast Precursor In Vitro

Bone remodeling is a complex process and it involves periods of deposition and resorption. Bone resorption is a process by which bone is broken down by osteoclasts in response to different stimuli. Osteoclast precursors differentiate into multinuclear osteoclasts in response to macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor Kappa-B ligand (RANKL). Under pathologic conditions, the cytokine profile is different and involves a mixture of inflammatory cytokines. Tumor necrosis factor alpha (TNF-&#945;) is one of the most important cytokines as it is found in large amounts in areas involved with inflammatory osteolysis. The purpose of this protocol is to provide a method by which murine bone marrow is isolated to generate osteoclasts through induction with M-CSF and either RANKL or TNF-&#945; which will be subsequently inhibited by increasing doses of anti-c-fms antibody, the receptor for M-CSF. This experiment highlights the therapeutic value of anti-c-fms antibody in diseases of inflammatory bone resorption.

This protocol includes dissection of murine long bones and bone marrow isolation, to generate bone marrow macrophages and produce osteoclasts using macrophage colony stimulating factor (M-CSF) and receptor activator of nuclear factor Kappa-B ligand (RANKL) or tumor necrosis factor alpha (TNF-&#945;) and their arrest by anti-c-fms antibody, M-CSF receptor.

Osteoclast 형성과 생체 외에서 Osteoclast 선구자의 확산에 안티-c-fms 항 체의 효과

Bone remodeling is a complex process and it involves periods of deposition and resorption. Bone resorption is a process by which bone is broken down by ...

면역학 및 감염병학

Osteoclast Derivation from Mouse Bone Marrow

Osteoclasts are highly specialized cells that are derived from the monocyte/macrophage lineage of the bone marrow. Their unique ability to resorb both the organic and inorganic matrices of bone means that they play a key role in regulating skeletal remodeling. Together, osteoblasts and osteoclasts are responsible for the dynamic coupling process that involves both bone resorption and bone formation acting together to maintain the normal skeleton during health and disease.
As the principal bone-resorbing cell in the body, changes in osteoclast differentiation or function can result in profound effects in the body. Diseases associated with altered osteoclast function can range in severity from lethal neonatal disease due to failure to form a marrow space for hematopoiesis, to more commonly observed pathologies such as osteoporosis, in which excessive osteoclastic bone resorption predisposes to fracture formation.
An ability to isolate osteoclasts in high numbers in vitro has allowed for significant advances in the understanding of the bone remodeling cycle and has paved the way for the discovery of novel therapeutic strategies that combat these diseases. 
Here, we describe a protocol to isolate and cultivate osteoclasts from mouse bone marrow that will yield large numbers of osteoclasts.

Osteoclasts are the principal bone-resorbing cell in the body. An ability to isolate osteoclasts in large numbers has resulted in significant advances in the understanding of osteoclast biology. In this protocol, we describe a method for isolation, cultivating and quantifying osteoclast activity in vitro.

마우스 골수에서 파골 세포의 유도

Osteoclasts are highly specialized cells that are derived from the monocyte/macrophage lineage of the bone marrow. Their unique ability to resorb both the ...

생물학

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

Mature osteoclasts are multinucleated cells that can degrade bone through the secretion of acids and enzymes. They play a crucial role in various diseases (e.g., osteoporosis and bone cancer) and are therefore important objects of research. In vitro, their activity can be analyzed by the formation of resorption pits. In this protocol, we describe a simple pit assay method using calcium phosphate (CaP) coated cell culture plates, which can be easily visualized and quantified. Osteoclast precursors derived from human peripheral blood mononuclear cells (PBMCs) were cultured on the coated plates in the presence of osteoclastogenic stimuli. After 9 days of incubation, osteoclasts were fixed and stained for fluorescence imaging while the CaP coating was counterstained by calcein. To quantify the resorbed area, the CaP coating on plates was stained with 5% AgNO3 and visualized by brightfield imaging. The resorption pit area was quantified using ImageJ.

A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro

Here, we present a simple and effective assay procedure for resorption pit assays using calcium phosphate coated cell culture plates.

시험관 내에서 파골세포 재흡수를 시각화하고 정량화하는 간단한 구덩이 분석 프로토콜

Mature osteoclasts are multinucleated cells that can degrade bone through the secretion of acids and enzymes. They play a crucial role in various diseases ...

Isolation, Purification, and Differentiation of Osteoclast Precursors from Rat Bone Marrow

Osteoclasts are large, multinucleated, and bone-resorbing cells of the monocyte-macrophage lineage that are formed by the fusion of monocytes or macrophage precursors. Excessive bone resorption is one the most significant cellular mechanisms leading to osteolytic diseases, including osteoporosis, periodontitis, and periprosthetic osteolysis. The main physiological function of osteoclasts is to absorb both the hydroxyapatite mineral component and the organic matrix of bone, generating the characteristic resorption appearance on the surface of bones. There are relatively few osteoclasts compared to other cells in the body, especially in adult bones. Recent studies have focused on how to obtain more mature osteoclasts in less time, which has always been a problem. Several improvements in the isolation and culture techniques have developed in laboratories in order to obtain more mature osteoclasts. Here, we introduce a method that isolates bone marrow in less time and with less effort compared to the traditional procedure, using a special and simple device. With the use of density gradient centrifugation, we obtain large amounts of fully differentiated osteoclasts from rat bone marrow, which are identified by classical methods.

Osteoclasts are tissue-specific macrophage polykaryons derived from the monocyte-macrophage lineage of hematopoietic stem cells. This protocol describes how to isolate bone marrow cells so that large quantities of osteoclasts are obtained while reducing the risk of accidents found in traditional methods.

쥐 골 골에서의 파 골 세포 전 구체의 분리, 정제 및 분화

Osteoclasts are large, multinucleated, and bone-resorbing cells of the monocyte-macrophage lineage that are formed by the fusion of monocytes or ...

Resorption Pit Assay: An In Vitro Technique to Quantify Calcium Resorption Activity of Mature Osteoclasts using Calcium Phosphate-Coated Culture Plates

Source: Cen, W., et al. <a href="https://www.jove.com/v/64016">A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro.</a> J. Vis. Exp. (2022)
In this video, we describe an in vitro osteoclastic resorption assay on calcium phosphate-coated cell culture plates cultured with human peripheral blood mononuclear cell-derived osteoclasts. The osteoclasts form a characteristic actin ring on the calcium phosphate-coated matrix of the cell culture plate and degrade the matrix within the actin ring region to release calcium and phosphate ions, resulting in the formation of distinct resorption pits within the degraded matrix.

Resorption Pit Assay: Calcium Phosphate-Coated Culture Plate를 사용하여 성숙한 골세포의 칼슘 흡수 활성을 정량화하는 In Vitro 기술

Source: Cen, W., et al. A Simple Pit Assay Protocol to Visualize and Quantify Osteoclastic Resorption In Vitro. J. Vis. Exp. (2022)
In this video, we ...

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Assay Techniques

Cell-based assays

Effect of the Anti C-fms Antibody on Osteoclastogenesis in Mouse Bone Marrow Cells In Vitro

Source: Marahleh, A., et al. <a href="https://app.jove.com/v/59089">Effect of Anti-c-fms Antibody on Osteoclast Formation and Proliferation of Osteoclast Precursor In Vitro.</a> J. Vis. Exp. (2019)
In this video, we demonstrate the effect of different anti-c-fms antibody concentrations on osteoclastogenesis, which is the process of osteoclast formation from osteoclast progenitors.

생쥐 골수 세포의 골수 형성에 대한 Anti C-fms 항체의 효과시험관 내

Source: Marahleh, A., et al. Effect of Anti-c-fms Antibody on Osteoclast Formation and Proliferation of Osteoclast Precursor In Vitro. J. Vis. Exp. 
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