대퇴골과 Tibiae에서 골수 수확 <ol><li> 12 주 된 마우스 (우리가 BALB / C 마우스를 사용하고 있지만 어떤 마우스 변형이 절차를 할 수있는)에 - 절차의 첫 단계는 6의 대퇴골과 tibiae에서 골수를 수확하는 것입니다. 시작하려면, CO 2 흡입 (표시되지 않습니다)로 마우스를 안락사. 각각의 뒷다리에서 수확 대퇴골과 tibiae. 그런 다음, 면도날과 고관절 및 무릎 관절을 제거하는 대퇴골의 각 끝을 잘라 골수를 노출합니다. </li><li> 비슷한 방식으로, 무릎 및 발목 인접 지역을 제거하고 골수를 노출 tibiae의 각 끝을 잘라. </li><li> 26 게이지 바늘로 3 ML의 주사기를 가지고 RPMI 10 % FBS, 페니실린, 스트렙토 마이신과 베타 - 메르 캅 토 에탄올 (BME = 100 음)와 보충으로 채우십시오. 핀셋을 사용하여 RPMI 미디어를 포함하는 페트리 접시 위에 뼈가시오. 뼈의 한쪽 끝을로 주사기 바늘을 삽입하여 골수를 제거하는 플런저를 우울. 주사기 바늘은 잔류 골수를 제거하는 뼈 내부 위아래로 이동할 수 있습니다. </li><li> 문화를 확립을 진행 후 모든 뼈에 대해 절차를 반복합니다. </li></ol> 문화 구축 <ol><li> 골수 문화를 구축하기 위해 세포 현탁액으로 조직을 해산시키기 위해 여러 번 다운 플러시 골수를 피펫합니다. </li><li> 50 ML 원뿔 튜브의 상단에 70 미크론 필터를 삽입하고, 필터에 세포 현탁액을 추가합니다. </li><li> 페트리 접시에게 RPMI 한 시간을 씻어 후 70 미크론 필터로 헹굼 추가합니다. </li><li> 1 ML 주사기 플런저의 고무 엔드를 사용하여 70 미크론 필터 위에 남아있는 골수 조각을 갈아서. </li><li> RPMI과 필터를 한 번 씻으십시오. </li><li> 필터를 세척 후 5 분 1,200 RPM의 세포 현탁액을 원심 분리기. </li><li> PBS에서 resuspending하여 세포 펠렛을 씻으십시오. </li><li> hemocytometer를 사용하여 셀을 계산합니다. 우리는 일반적으로 마우스를 한번의 유골 (두 대퇴골 두 개의 tibiae)에서 ~ 90000000 세포를 구하십시오. 세포를 계산하면 5 분 동안 1,200 rpm으로 그들을 다시 원심 분리기. </li><li> RPMI 및 나누어지는 그 때문에 최종 농도가 ML 당 백만 세포는 충분한 RPMI를 포함하는 조직 문화 flasks에 소량의 세포를 Resuspend. </li><li> 관심의 세포 유형의 개발을 촉진하기 크린 시토킨를 추가합니다. 이 경우 시토킨 인터루킨 - 3 (IL - 3 = 10 NG / ML)은 basophils과 마스트 세포의 발전을 촉진하기 위해 추가됩니다. basophils를 얻기위한 10 일 동안 알을 품다. 돛대 세포의 경우, 5 주 동안 알을 품다. 돛대 세포를 생성할 때, 미디어와 IL - 3는 일주일에 한 번씩 변경해야합니다. </li><li> 여기 마스트 세포가 단지 도금 후 나타나는 방법을 좀 봐입니다. 이 이미지는 10 일 인터루킨 - 3의 추가 후 basophils과 마스트 세포로 마스트 골수 세포의 분화를 보여줍니다. </li><li> 원하는 세포 유형이 취득하면, electroporation 단계로 진행하십시오. </li></ol> Electroporating 셀 <ol><li> electroporation 단계를 시작하려면 문화 플라스크에 세포의 수를 결정합니다. </li><li> 세포는 일반적으로 ML 당 10 만명의 밀도에 electroporated되므로, 원뿔 튜브에 필요한 휴대폰 번호를 전송하고 5 분 1,200 RPM에서 튜브를 원심 분리기. </li><li> centrifuging 후, 미디어를 대기음 1X PBS로 세포를 씻어, 5 분 동안 1,200 rpm으로 다시 원심 분리기. </li><li> PBS를 기음과 ML 당 10,000,000 세포의 세포 현탁액을 만드는 바이오 래드 진 Pulser의 electroporation 버퍼를 추가합니다. </li><li> 세포를 resuspending 후, ML 당 10-20 마이크로 그램의 최종 농도에서 원하는 플라스미드 DNA를 추가합니다. </li><li> 96 - 웰 플레이트 electroporation에 선택의 우물에 세포 현탁액의 나누어지는 150 microliters. </li><li> MXcell 플레이트 챔버의 electroporation 판을 넣고 뚜껑을 닫습니다. </li><li> 돛대 세포의 transfection하기 전에, electroporation 프로토콜은 미리 또는 저장 프로토콜을 사용하지 않는 MXcell로 프로그래밍해야합니다. 최적화 실험을 통해 우리는 마스트 세포의 가장 높은 transfection 효율이 사각형 웨이브 펄스 프로토콜을 사용하여 발생하는 것을 발견했습니다. 우리는 2000uF 1000 ohms에서 300V/20ms, 350/15ms, 350V/20ms 및 350V/10ms 스퀘어 웨이브 펄스를 제공하기 위해 접시에 electroporation 조건을 달라질 수 있습니다. 프로토콜 설정하고 장치에 로드된되면, 세포를 electroporate에 &quot;펄스&quot;를 누르십시오. </li><li> electroporation가 완료되면, 조직 문화 접시에 세포를 전송합니다. 우리는 일반적으로 ML 인터루킨 - 3 당 10 나노 그램 300 microliters RPMI를 (10 % FCS, 페니실린, 스트렙토 마이신과 베타 - 메르 캅 토 에탄올과 보충)가 포함된 48 잘 조직 배양 플레이트에 잘 각 150 microliter의 electroporation 샘플을 전송할 수 있습니다. 전지는 37 하룻밤 incubated 아르 ° C, 그때 표현과 억제를 위해 24 시간 후에 assayed. </li></ol> 트랜sfection 결과 플라스미드 ML 당 20 마이크로 그램 GFP를 사용하여 성공적인 electroporation 후 세포의 형광 현미경 이미지는 동영상에 표시됩니다. 약 30 %의 MXcell의 electroporation 시스템 transfection 효율성을 사용하는 것은 얻을 수 있습니다. 이 시스템은 세포 생존을 유지하면서 당신이 당신의 transfection 효율을 극대화하기 위해 조건을 변경할 수 있습니다.

더 많은 게놈 정보를 얻습니다 siRNA와 같은 새로운 도구를 개발로서, 생리학 관련 세포의 사용은 질병 경로, 단백질 - 단백질 상호 작용 및 신호 전달에 대한 우리의 이해를 심화하기 위해 점점 더 중요 해지고있다. 차 전지는 조직이나 체액에서 직접 얻은하고 체외에서 재배하고 있습니다. 이러한 세포는 외인성 유전 물질의 도입을 포함 방법의 숫자 조작하실 수 있습니다. 그것은 기본 세포에 플라스미드 DNA 또는 siRNA을 소개하는 가장 효과적인 방법은 electroporation 것을 점점 더 분명 해지고 있습니다. Electroporation은 transiently 따라서 외인성 핵산은 세포를 입력할 수 있도록 세포막의 투과성을 증가하기 위해 전기 펄스로 세포를 제공합니다. transfection의이 방법은 다른 세포 유형 / 라인, 따라서, 돛대 세포를 transfect하는 데 사용할 수있는뿐만 아니라 다른 골수 - 유래 세포 사이의 차이에 대한 수용하도록 최적화할 수 있습니다. 또한, 바이러스 - 매개 transfection와는 달리, electroporation은 transfected DNA의 크기에 제한을 부과하거나 광범위한 준비가 필요하지 않습니다. 지질 - 중재 transfection 대​​조적으로, electroporation 덜 독성 수 있으며 transfected 핵산의 endosomal 트래핑 발생하지 않습니다. 바이오 래드는 이러한 세포를 위해 특별히 MXcell의 electroporation 시스템을 개발했습니다. MXcell 여러분 transfection의 효율성과 세포 생존을 최대화하기 위해 조건을 변경할 수 있습니다. transfection 효율을 최적화하고 세포 사망, 전압을 포함하여 electroporation 매개 변수의 다수를 최소화하기 위해, 정전 용량, 저항, 및 펄스 길이는 조정과 평가 수 있습니다.

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<td>Gene Pulser MXcell Electroporation System</td>
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<td>Gene Pulser Electroporation Buffer</td>
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the preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation

그것은 electroporation은 기본 세포에 플라스미드 DNA 또는 siRNA을 소개하는 가장 효과적인 방법임을 더욱 분명 해지고 있습니다. 진 Pulser MXcell의 electroporation 시스템과 진 Pulser의 electroporation 버퍼가 특히 포유 동물 세포와 같은 기본 및 줄기 cells.This 비디오와 같은 어려운 transfect 세포로 핵산을 transfect하기 위해 개발되었습니다 것은 murine 골수에서 기본 조혈 세포 문화를 확립하는 방법을 보여줍니다 다음 MXcell 시스템의 electroporation을 위해 그들을 준비합니다. 우리는 대퇴골과 경골을 분리하여 시작합니다. 대퇴골과 경골 모두에서 골수 그런 다음 수확하고 문화가 설립됩니다. 교양 골수 세포는 다음 transfected 및 분석입니다.

Watch this Scientific Journal Video about The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation at JoVE.com

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

이 절차는 murine 골수에서 조혈 세포 기본 문화를 확립하고 진 Pulser MXCell의 electroporation 시스템을 사용 transfection하여 다음 방법을 설명합니다.

Electroporation에 대한 Murine 골수에서 조혈 세포 차 문화의 준비

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser ...

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25.6K 조회수.  Bio-Rad Laboratories, Inc.  ChineseVersion 

비디오: The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

Derivation of Hematopoietic Stem Cells from Murine Embryonic Stem Cells

A stem cell is defined as a cell with the capacity to both self-renew and generate multiple differentiated progeny.  Embryonic stem cells (ESC) are derived from the blastocyst of the early embryo and are pluripotent in differentiative ability.  Their vast differentiative potential has made them the focus of much research centered on deducing how to coax them to generate clinically useful cell types.  The successful derivation of hematopoietic stem cells (HSC) from mouse ESC has recently been accomplished and can be visualized in this video protocol.  HSC, arguably the most clinically exploited cell population, are used to treat a myriad of hematopoietic malignancies and disorders.  However, many patients that might benefit from HSC therapy lack access to suitable donors.  ESC could provide an alternative source of HSC for these patients.  The following protocol establishes a baseline from which ESC-HSC can be studied and inform efforts to isolate HSC from human ESC.  In this protocol, ESC are differentiated as embryoid bodies (EBs) for 6 days in commercially available serum pre-screened for optimal hematopoietic differentiation.  EBs are then dissociated and infected with retroviral HoxB4.  Infected EB-derived cells are plated on OP9 stroma, a bone marrow stromal cell line derived from the calvaria of  M-CSF-/- mice, and co-cultured in the presence of hematopoiesis promoting cytokines for ten days.  During this co-culture, the infected cells expand greatly, resulting in the generation a heterogeneous pool of 100s of millions of cells.  These cells can then be used to rescue and reconstitute lethally irradiated mice.

This protocol details the derivation of transplantable hematopoietic stem cells from mouse embryonic stem cells (ESC) and their subsequent injection into lethally irradiated recipient mice. Briefly, ESC are differentiated as embryoid bodies, which are then infected with retroviral HoxB4 and co-cultured with OP9 stromal cells and hematopoietic cytokines.

Murine 배아 줄기 세포로부터 조혈 줄기 세포의 유도

A stem cell is defined as a cell with the capacity to both self-renew and generate multiple differentiated progeny.  Embryonic stem cells (ESC) are ...

연구

생물학

Primary Neuronal Cultures from the Brains of Late Stage Drosophila Pupae

In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. The procedure begins with the removal of brains from animals at 70-78 hrs after puparium formation. The isolated brains are shown after brief incubation in papain followed by several washes in serum-free growth medium. The process of mechanical dissociation of each brain in a 5 ul drop of media on a coverslip is illustrated. The axons and dendrites of the post-mitotic neurons are sheered off near the soma during dissociation but the neurons begin to regenerate processes within a few hours of plating. Images show live cultures at 2 days. Neurons continue to elaborate processes during the first week in culture. Specific neuronal populations can be identified in culture using GAL4 lines to drive tissue specific expression of fluorescent markers such as  GFP or RFP. Whole cell recordings have demonstrated the cultured neurons form functional, spontaneously active cholinergic and GABAergic synapses. A short video segment illustrates calcium dynamics in the cultured neurons using Fura-2 as a calcium indicator dye to monitor spontaneous calcium transients and nicotine evoked calcium responses in a dish of cultured neurons. These pupal brain cultures are a useful model system in which genetic and pharmacological tools can be used to identify intrinsic and extrinsic factors that influence formation and function of central synapses.

This video demonstrates the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. Views of live cultures show neurite outgrowth and imaging of calcium levels using Fura-2.

늦은 단계 Drosophila Pupae의 두뇌에서 기본의 연결을 문화

In this video, we demonstrate the preparation of primary neuronal cultures from the brains of late stage Drosophila pupae. The procedure begins with the ...

Electroporation of Mycobacteria

High efficiency transformation is a major limitation in the study of mycobacteria. The genus Mycobacterium can be difficult to transform; this is mainly caused by the thick and waxy cell wall, but is compounded by the fact that most molecular techniques have been developed for distantly-related species such as Escherichia coli and Bacillus subtilis. In spite of these obstacles, mycobacterial plasmids have been identified and DNA transformation of many mycobacterial species have now been described. The most successful method for introducing DNA into mycobacteria is electroporation. Many parameters contribute to successful transformation; these include the species/strain, the nature of the transforming DNA, the selectable marker used, the growth medium, and the  conditions for the electroporation pulse. Optimized methods for the transformation of both slow- and fast-grower are detailed here. Transformation efficiencies for different mycobacterial species and with various selectable markers are reported.

Mycobacterial pathogenic strategies remain poorly understood. The slow growth rate of most species, the impenetrable nature of the cell-wall, and the hazards of working with pathogens make mycobacteria difficult to study and are largely responsible for our poor understanding of these organisms. In this video we will demonstrate the technique of electroporation, which involves subjecting cells to a brief high electrical impulse to allow the entry of DNA. It is the most widely used method for introducing DNA into mycobacterial cells.

Mycobacteria의 Electroporation

High efficiency transformation is a major limitation in the study of mycobacteria. The genus Mycobacterium can be difficult to transform; this is mainly ...

Culture of myeloid dendritic cells from bone marrow precursors

Myeloid dendritic cells (DCs) are frequently used to study the interactions between innate and adaptive immune mechanisms and the early response to infection. Because these are the most potent antigen presenting cells, DCs are being increasingly used as a vaccine vector to study the induction of antigen-specific immune responses. In this video, we demonstrate the procedure for harvesting tibias and femurs from a donor mouse, processing the bone marrow and differentiating DCs in vitro. The properties of DCs change following stimulation: immature dendritic cells are potent phagocytes, whereas mature DCs are capable of antigen presentation and interaction with CD4+ and CD8+ T cells. This change in functional activity corresponds with the upregulation of cell surface markers and cytokine production. Many agents can be used to mature DCs, including cytokines and toll-like receptor ligands. In this video, we demonstrate flow cytometric comparisons of expression of two co-stimulatory molecules, CD86 and CD40, and the cytokine, IL-12, following overnight stimulation with CpG or mock treatment. After differentiation, DCs can be further manipulated for use as a vaccine vector or to generate antigen-specific immune responses by in vitro pulsing using peptides or proteins, or transduced using recombinant viral vectors. 

This video demonstrates the procedure for differentiating myeloid dendritic cells from mouse bone marrow. Isolation of mouse tibia and femur, and processing of bone marrow are demonstrated. Pictures demonstrating cell morphology before and after differentiation, and figures depicting cell phenotype and IL-12 production following maturation using CpG are shown.

골수 엽 성의 전구 물질에서 골수양 돌기 세포의 문화

Myeloid dendritic cells (DCs) are frequently used to study the interactions between innate and adaptive immune mechanisms and the early response to ...

Primary Dissociated Midbrain Dopamine Cell Cultures from Rodent Neonates

The ability to create primary cell cultures of dopamine neurons allows for the study of the presynaptic characteristics of dopamine neurons in isolation from systemic input from elsewhere in the brain. In our lab, we use these neurons to assess dopamine release kinetics using carbon fiber amperometry, as well as expression levels of dopamine related genes and proteins using quantitative PCR and immunocytochemistry. In this video, we show you how we generate these cultures from rodent neonates.
The process involves several steps, including the plating of cortical glial astrocytes, the conditioning of neuronal cell culture media by the glial substrate, the dissection of the midbrain in neonates, the digestion, extraction and plating of dopamine neurons and the addition of neurotrophic factors to ensure cell survival.
The applications suitable for such a preparation include electrophysiology, immunocytochemistry, quantitative PCR, video microscopy (i.e., of real-time vesicular fusion with the plasma membrane), cell viability assays and other toxicological screens.

Primary dissociated midbrain dopamine cell cultures allow for the study of presynaptic characteristics of dopamine neurons. They can be used to monitor real-time dopamine release kinetics and protein/mRNA levels of regulators of dopamine exocytosis. Here, we show you how to generate these cultures from rodent neonates.

쥐 Neonates에서 기본 Dissociated Midbrain의 도파민 세포 문화

The ability to create primary cell cultures of dopamine neurons allows for the study of the presynaptic characteristics of dopamine neurons in isolation ...

Homing of Hematopoietic Cells to the Bone Marrow

Homing is the phenomenon whereby transplanted hematopoietic cells are able to travel to and engraft or establish residence in the bone marrow. Various chemomkines and receptors are involved in the homing of hematopoietic stem cells. [1, 2]
This paper outlines the classic homing protocol used in hematopoietic stem cell studies. In general this involves isolating the cell population whose homing needs to be investigated, staining this population with a dye of interest and injecting these cells into the blood stream of a recipient animal. The recipient animal is then sacrificed at a pre-determined time after injection and the bone marrow evaluated for the percentage or absolute number of cells which are positive for the dye of interest. In one of the most common experimental schemes, the homing efficiency of hematopoietic cells from two genetically distinct animals (a wild type animal and the corresponding knock-out) is compared. This article describes the hematopoietic cell homing protocol in the framework of such as experiment.

This article describes a protocol used to study the homing of hematopoietic cells to their niches in the bone marrow.

골수에 조혈 세포의 유도

Homing is the phenomenon whereby transplanted hematopoietic cells are able to travel to and engraft or establish residence in the bone marrow.  Various ...

Primary Culture and Plasmid Electroporation of the Murine Organ of Corti. 

In all mammals, the sensory epithelium for audition is located along the spiraling organ of Corti that resides within the conch shaped cochlea of the inner ear (fig 1). Hair cells in the developing cochlea, which are the mechanosensory cells of the auditory system, are aligned in one row of inner hair cells and three (in the base and mid-turns) to four (in the apical turn) rows of outer hair cells that span the length of the organ of Corti. Hair cells transduce sound-induced mechanical vibrations of the basilar membrane into neural impulses that the brain can interpret. Most cases of sensorineural hearing loss are caused by death or dysfunction of cochlear hair cells.
An increasingly essential tool in auditory research is the isolation and in vitro culture of the organ explant 1,2,9. Once isolated, the explants may be utilized in several ways to provide information regarding normative, anomalous, or therapeutic physiology. Gene expression, stereocilia motility, cell and molecular biology, as well as biological approaches for hair cell regeneration are examples of experimental applications of organ of Corti explants.
This protocol describes a method for the isolation and culture of the organ of Corti from neonatal mice. The accompanying video includes stepwise directions for the isolation of the temporal bone from mouse pups, and subsequent isolation of the cochlea, spiral ligament, and organ of Corti. Once isolated, the sensory epithelium can be plated and cultured in vitro in its entirety, or as a further dissected micro-isolate that lacks the spiral limbus and spiral ganglion neurons. Using this method, primary explants can be maintained for 7-10 days. As an example of the utility of this procedure, organ of Corti explants will be electroporated with an exogenous DsRed reporter gene. This method provides an improvement over other published methods because it provides reproducible, unambiguous, and stepwise directions for the isolation, microdissection, and primary culture of the organ of Corti.

Primary Culture and Plasmid Electroporation of the Murine Organ of Corti.

This procedure describes a method for the isolation and culture of the murine organ of Corti with or without the spiral limbus and spiral ganglion neurons. We also demonstrate a method for the expression of an exogenous reporter gene in the organ of Corti explant by electroporation.

차 문화와 Corti의 Murine 오르간의 플라스미드 Electroporation.

In all mammals, the sensory epithelium for audition is located along the spiraling organ of Corti that resides within the conch shaped cochlea of the ...

Primary Cell Cultures from Drosophila Gastrula Embryos

Here we describe a method for preparing and culturing primary cells dissociated from Drosophila gastrula embryos. In brief, a large amount of staged embryos from young and healthy flies are collected, sterilized, and then physically dissociated into a single cell suspension using a glass homogenizer. After being plated on culture plates or chamber slides at an appropriate density in culture medium, these cells can further differentiate into several morphologically-distinct cell types, which can be identified by their specific cell markers. Furthermore, we present conditions for treating these cells with double stranded (ds) RNAs to elicit gene knockdown. Efficient RNAi in Drosophila primary cells is accomplished by simply bathing the cells in dsRNA-containing culture medium. The ability to carry out effective RNAi perturbation, together with other molecular, biochemical, cell imaging analyses, will allow a variety of questions to be answered in Drosophila primary cells, especially those related to differentiated muscle and neuronal cells.

Primary Cell Cultures from Drosophila Gastrula Embryos

We provide a detailed protocol for preparing primary cells dissociated from Drosophila embryos. The ability to carry out the effective RNAi perturbation, together with other molecular, biochemical and cell imaging methods will allow a variety of questions to be addressed in Drosophila primary cells.

에서 기본 셀 문화 Drosophila Gastrula의 태아

Here we describe a method for preparing and culturing primary cells dissociated from Drosophila gastrula embryos.   In brief, a large amount of staged ...

Preparation of Primary Myogenic Precursor Cell/Myoblast Cultures from Basal Vertebrate Lineages

Due to the inherent difficulty and time involved with studying the myogenic program in vivo, primary culture systems derived from the resident adult stem cells of skeletal muscle, the myogenic precursor cells (MPCs), have proven indispensible to our understanding of mammalian skeletal muscle development and growth. Particularly among the basal taxa of Vertebrata, however, data are limited describing the molecular mechanisms controlling the self-renewal, proliferation, and differentiation of MPCs. Of particular interest are potential mechanisms that underlie the ability of basal vertebrates to undergo considerable postlarval skeletal myofiber hyperplasia (i.e.&#160;teleost fish) and full regeneration following appendage loss (i.e.&#160;urodele amphibians). Additionally, the use of cultured myoblasts could aid in the understanding of regeneration and the recapitulation of the myogenic program and the differences between them. To this end, we describe in detail a robust and efficient protocol (and variations therein) for isolating and maintaining MPCs and their progeny, myoblasts and immature myotubes, in cell culture as a platform for understanding the evolution of the myogenic program, beginning with the more basal vertebrates. Capitalizing on the model organism status of the zebrafish (Danio rerio), we report on the application of this protocol to small fishes of the cyprinid clade Danioninae. In tandem, this protocol can be utilized to realize a broader comparative approach by isolating MPCs from the Mexican axolotl (Ambystomamexicanum) and even laboratory rodents. This protocol is now widely used in studying myogenesis in several fish species, including rainbow trout, salmon, and sea bream1-4.

In vitro culture systems have proven indispensible to our understanding of vertebrate myogenesis. However, much remains to be learned about nonmammalian skeletal muscle development and growth, particularly in basal taxa. An efficient and robust protocol for isolating the adult stem cells of this tissue, the myogenic precursor cells (MPCs), and maintaining their self-renewal, proliferation, and differentiation in a primary culture setting allows for the identification of conserved and divergent regulatory mechanisms throughout the vertebrate lineages.

기초 척추 계통의 주요 근원 성 전구 세포 / 근원 세포 배양의 준비

Due to the inherent difficulty and time involved with studying the myogenic program in vivo, primary culture systems derived from the resident adult stem ...

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 ...