Name: the preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation
Uploaded: 2009-01-06T23:59:00.000+00:00
Duration: 8 min 15 s
Description: Watch this Scientific Journal Video about The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation at JoVE.com

La raccolta di midollo osseo da femori e tibie <ol><li> Il primo passo della procedura è quello di raccogliere midollo osseo dal femore e la tibia di un 6 - a 12 settimane vecchio mouse (stiamo usando topi BALB / c, ma si può fare questa procedura con qualsiasi ceppo di topi). Per iniziare, eutanasia il mouse da CO 2 inalazioni (non sarà mostrato). Vendemmia femore e tibia dalle zampe posteriori di ciascuno. Poi, con una lametta da barba, tagliare ogni estremità del femori per rimuovere l&#39;anca e le articolazioni del ginocchio e per esporre il midollo. </li><li> In modo simile, tagliare ogni estremità della tibia per rimuovere il ginocchio e della caviglia regioni limitrofe e per esporre il midollo. </li><li> Fate una siringa da 3 ml con una 26-gauge e riempirlo con RPMI supplementato con 10% FBS, penicillina, streptomicina, e beta-mercaptoetanolo (BME = 100 uM). Utilizzando una pinzetta, tenere l&#39;osso su una piastra di Petri contenente RPMI media. Inserire l&#39;ago della siringa in una delle estremità delle ossa e premere lo stantuffo per scovare il midollo osseo. L&#39;ago della siringa può essere spostato su e giù dentro l&#39;osso per scovare midollo residuo. </li><li> Ripetere la procedura per tutte le ossa, e quindi procedere con istituisce la culture. </li></ol> Stabilire Culture <ol><li> Per stabilire le culture del midollo osseo, pipetta il midollo osseo lavata su e giù parecchie volte per rompere il tessuto in una sospensione cellulare. </li><li> Collocare un filtro di 70 micron in cima ad un tubo 50 ml conica, e aggiungere la sospensione cellulare al filtro. </li><li> Sciacquare la piastra Petri una volta con RPMI, quindi aggiungere il risciacquo del filtro per i 70 micron. </li><li> Utilizzando la fine di gomma di una siringa da 1 ml stantuffo, macinare i pezzi rimanenti del midollo osseo nella parte superiore del filtro di 70 micron. </li><li> Lavare il filtro una volta con RPMI. </li><li> Dopo aver lavato il filtro, centrifuga la sospensione cellulare a 1200 rpm per 5 minuti. </li><li> Lavare il pellet da risospendere in PBS. </li><li> Contare le cellule utilizzando un emocitometro. Noi di solito ottenere ~ 90 milioni di cellule di ossa di un mouse (due femori e due tibie). Dopo il conteggio delle cellule, li centrifuga di nuovo a 1200 rpm per 5 minuti. </li><li> Risospendere le celle di una piccola quantità di RPMI e le aliquote in fiasche colture di tessuti contenenti abbastanza RPMI in modo che l&#39;concentrazione finale è di 1 milione di cellule per mL. </li><li> Aggiungi citochine a promuovere lo sviluppo del vostro tipo di cellula di interesse. In questo caso, la citochina interleuchina-3 (IL-3 = 10 ng / mL) viene aggiunto per promuovere lo sviluppo dei basofili e mastociti. Per acquisire basofili, incubare per 10 giorni. Per i mastociti, incubare per 5 settimane. Quando si genera mastociti, i media e IL-3 dovrebbe essere cambiata una volta alla settimana. </li><li> Ecco uno sguardo a come mastociti dovrebbe apparire subito dopo placcatura. Questa immagine mostra la differenziazione delle cellule del midollo dell&#39;albero in basofili e mastociti 10 giorni dopo l&#39;aggiunta di interleuchina-3. </li><li> Una volta che il tipo cellulare desiderato si ottiene, procedere con il passaggio elettroporazione. </li></ol> Cellule Electroporating <ol><li> Per iniziare il passo elettroporazione, determinare il numero di cellule nel pallone cultura. </li><li> Le cellule sono in genere elettroporate ad una densità di 10 milioni per ml, in modo da trasferire il numero di cellule per tubi conici e centrifugare le provette a 1200 rpm per 5 minuti. </li><li> Dopo la centrifugazione, aspirare i media, lavare le cellule con PBS 1X, e centrifugare di nuovo a 1200 rpm per 5 minuti. </li><li> Aspirare il PBS e aggiungere Bio-Rad Gene buffer di elettroporazione Pulser per fare una sospensione di cellule di 10 milioni di cellule per ml. </li><li> Dopo risospendere le cellule, aggiungere il DNA desiderato plasmide ad una concentrazione finale di 10 a 20 microgrammi per ml. </li><li> Aliquota 150 microlitri di sospensione cellulare in pozzi di vostra scelta su un piatto elettroporazione 96-bene. </li><li> Mettete la placca nella camera di elettroporazione MXcell piastra e chiudere il coperchio. </li><li> Prima di trasfezione dei mastociti, un protocollo di elettroporazione deve essere programmato in MXcell a meno di utilizzare un protocollo predefinito o memorizzati. Attraverso esperimenti di ottimizzazione che hanno scoperto che la più alta efficienza di trasfezione dei mastociti avvenire utilizzando protocolli di piazza dell&#39;onda di polso. Noi variare le condizioni elettroporazione sul piatto per fornire 300V/20ms, 350/15ms, 350V/20ms e 350V/10ms impulsi a onda quadra 2000uF e 1000 ohm. Una volta che il protocollo è stato impostato e caricata sul dispositivo, premere il tasto &quot;Pulse&quot; per electroporate le cellule. </li><li> Dopo l&#39;elettroporazione è completa, trasferire le cellule a una piastra di coltura dei tessuti. Noi di solito trasferire ogni campione elettroporazione 150 microlitri di un pozzo in un 48-pozzetti di coltura contenente 300 microlitri RPMI (supplementato con 10% FCS, penicillina, streptomicina, e beta-mercaptoetanolo) con 10 nanogrammi per ml interleuchina-3. Le cellule vengono incubate overnight a 37 ° C, poi analizzati 24 ore più tardi per l&#39;espressione e la soppressione. </li></ol> Transfection Risultati L&#39;immagine di microscopia a fluorescenza delle cellule dopo l&#39;elettroporazione di successo con 20 microgrammi per ml GFP plasmide viene mostrato nel video. Utilizzando il sistema di elettroporazione MXcell efficienze di trasfezione di circa il 30% può essere ottenuto. Il sistema consente di variare le condizioni per massimizzare l&#39;efficienza di trasfezione, pur mantenendo la vitalità cellulare.

Come più informazioni genomiche emerge e nuovi strumenti come siRNA sono sviluppati, l&#39;uso di cellule fisiologicamente rilevanti è diventato sempre più importante per migliorare la nostra comprensione delle vie di malattia, interazioni proteina-proteina, e la trasduzione del segnale. Pile sono ottenute direttamente da tessuti o fluidi e coltivate in vitro. Queste cellule possono essere manipolati in vari modi, anche attraverso l&#39;introduzione di materiale genetico esogeno. E &#39;sempre più evidente che il modo più efficace per introdurre DNA plasmidico o siRNA in cellule primarie è elettroporazione. Elettroporazione espone le cellule a impulsi elettrici al fine di aumentare temporaneamente la permeabilità della membrana cellulare, consentendo in tal modo esogeno acidi nucleici per entrare nella cellula. Questo metodo di trasfezione può essere ottimizzato per accogliere le differenze tra diversi tipi di cellule / linee e, quindi, può essere utilizzato per trasfettare mastociti così come qualsiasi altro osso cellule derivate dal midollo. Inoltre, a differenza virale mediata trasfezione, elettroporazione non impone limiti alle dimensioni del DNA transfettate, né richiedono una preparazione. A differenza dei lipidi mediata trasfezione, elettroporazione può essere meno tossico e non dia luogo a cattura endosomiali degli acidi nucleici transfettate. Bio-Rad ha sviluppato il sistema di elettroporazione MXcell appositamente per queste cellule. il MXcell consente di variare le condizioni per massimizzare l&#39;efficienza di trasfezione e la vitalità cellulare. Al fine di ottimizzare l&#39;efficienza di trasfezione e ridurre al minimo la morte delle cellule, una moltitudine di parametri tra cui elettroporazione tensione, capacità, resistenza e durata dell&#39;impulso può essere regolata e valutati.

<table><tbody><tr><td>Gene Pulser MXcell Electroporation System</td><td>Bio-Rad</td><td>165-2670</td><td></td></tr><tr><td>Gene Pulser Electroporation Buffer</td><td>Bio-Rad</td><td>165-2677</td><td></td></tr></tbody></table>

the preparation of primary hematopoietic cell cultures from murine bone marrow for electroporation

E &#39;sempre più evidente che elettroporazione è il modo più efficace per introdurre DNA plasmidico o siRNA in cellule primarie. Il gene Pulser sistema di elettroporazione MXcell e Gene buffer di elettroporazione Pulser sono stati specificamente sviluppati per trasfettare acidi nucleici nelle cellule dei mammiferi e di difficile trasfezione delle cellule, come i video cells.This primarie e staminali dimostra come stabilire colture primarie di cellule emopoietiche dal midollo osseo murino , e poi li preparano per l&#39;elettroporazione nel sistema MXcell. Si comincia isolando femore e della tibia. Midollo osseo da femore e della tibia vengono poi raccolte e le culture sono stabiliti. Colture di cellule del midollo osseo sono poi transfettate e analizzati.

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

Questa procedura descrive come stabilire colture primarie di cellule emopoietiche dal midollo osseo murino ed è seguita dalla trasfezione con il gene Pulser sistema di elettroporazione MXCell.

La preparazione di colture primarie di cellule emopoietiche dal midollo osseo murino per elettroporazione

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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JoVE Journal

Biology

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

Highly Efficient Gene Disruption of Murine and Human Hematopoietic Progenitor Cells by CRISPR/Cas9

Advances in the hematopoietic stem cell (HSCs) field have been aided by methods to genetically engineer primary progenitor cells as well as animal models. Complete gene ablation in HSCs required the generation of knockout mice from which HSCs could be isolated, and gene ablation in primary human HSCs was not possible. Viral transduction could be used for knock-down approaches, but these suffered from variable efficacy. In general, genetic manipulation of human and mouse hematopoietic cells was hampered by low efficiencies and extensive time and cost commitments. Recently, CRISPR/Cas9 has dramatically expanded the ability to engineer the DNA of mammalian cells. However, the application of CRISPR/Cas9 to hematopoietic cells has been challenging, mainly due to their low transfection efficiencies, the toxicity of plasmid-based approaches and the slow turnaround time of virus-based protocols.
A rapid method to perform CRISPR/Cas9-mediated gene editing in murine and human hematopoietic stem and progenitor cells with knockout efficiencies of up to 90% is provided in this article. This approach utilizes a ribonucleoprotein (RNP) delivery strategy with a streamlined three-day workflow. The use of Cas9-sgRNA RNP allows for a hit-and-run approach, introducing no exogenous DNA sequences in the genome of edited cells and reducing off-target effects. The RNP-based method is fast and straightforward: it does not require cloning of sgRNAs, virus preparation or specific sgRNA chemical modification. With this protocol, scientists should be able to successfully generate knockouts of a gene of interest in primary hematopoietic cells within a week, including downtimes for oligonucleotide synthesis. This approach will allow a much broader group of users to adapt this protocol for their needs.

A protocol for fast CRISPR/Cas9-mediated gene disruption in mouse and human primary hematopoietic cells is described in this article. Cas9-sgRNA ribonucleoproteins are introduced via electroporation with sgRNAs generated through in vitro transcription and commercial Cas9. High editing efficiencies are achieved with limited time and financial cost.

Rottura del Gene altamente efficiente di cellule progenitrici ematopoietiche Murine ed umane di CRISPR/Cas9

Advances in the hematopoietic stem cell (HSCs) field have been aided by methods to genetically engineer primary progenitor cells as well as animal models. ...

Ricerca

Genetica

Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

As a subtype of leukocytes and progenitors of macrophages, monocytes are involved in many important processes of organisms and are often the subject of various fields in biomedical science. The method described below is a simple and effective way to isolate murine monocytes from heterogeneous bone marrow.
Bone marrow from the femur and tibia of Balb/c mice is harvested by flushing with phosphate buffered saline (PBS). Cell suspension is supplemented with macrophage-colony stimulating factor (M-CSF) and cultured on ultra-low attachment surfaces to avoid adhesion-triggered differentiation of monocytes. The properties and differentiation of monocytes are characterized at various intervals. Fluorescence activated cell sorting (FACS), with markers like CD11b, CD115, and F4/80, is used for phenotyping. At the end of cultivation, the suspension consists of 45%&#177; 12% monocytes. By removing adhesive macrophages, the purity can be raised up to 86%&#177; 6%. After the isolation, monocytes can be utilized in various ways, and one of the most effective and common methods for in vivo delivery is intravenous tail vein injection. 
This technique of isolation and application is important for mouse model studies, especially in the fields of inflammation or immunology. Monocytes can also be used therapeutically in mouse disease models.

Here we present a protocol that generates large amounts of murine monocytes from heterogeneous bone marrow for translational applications. In comparison to others, this new method helps reduce the number of sacrificed animals and lowers costs by avoiding expensive methods such as high gradient magnetic cell separation (MACS).

Isolamento e iniezione endovenosa di Murine derivate dal midollo osseo monociti

As a subtype of leukocytes and progenitors of macrophages, monocytes are involved in many important processes of organisms and are often the subject of ...

Immunologia e infezioni

Using the Gene Pulser MXcell Electroporation System to Transfect Primary Cells with High Efficiency

It is becoming increasingly apparent that electroporation is the most effective way to introduce plasmid DNA or siRNA into primary cells. The Gene Pulser MXcell electroporation system and Gene Pulser electroporation buffer (Bio-Rad) were specifically developed to easily transfect nucleic acids into mammalian cells and difficult-to-transfect cells, such as primary and stem cells. We will demonstrate how to perform a simple experiment to quickly identify the best electroporation conditions. We will demonstrate how to run several samples through a range of electroporation conditions so that an experiment can be conducted at the same time as optimization is performed. We will also show how optimal conditions identified using 96-well electroporation plates can be used with standard electroporation cuvettes, facilitating the switch from electroporation plates to electroporation cuvettes while maintaining the same electroporation efficiency. In the video, we will also discuss some of the key factors that can lead to the success or failure of electroporation experiments.

This procedure shows how to use the Gene Pulser MXcell electroporation system to rapidly and easily identify the best electroporation conditions for mouse embryonic fibroblasts (MEFs) or other primary cells. Considerations for troubleshooting are also discussed in the associated video.

Utilizzando il sistema di elettroporazione Gene Pulser MXcell per trasfettare cellule primarie con alta efficienza

Biologia

Phenotypic Analysis and Isolation of Murine Hematopoietic Stem Cells and Lineage-committed Progenitors

The bone marrow is the principal site where HSCs and more mature blood cells lineage progenitors reside and differentiate in an adult organism. HSCs constitute a minute cell population of pluripotent cells capable of generating all blood cell lineages for a life-time1. The molecular dissection of HSCs homeostasis in the bone marrow has important implications in hematopoiesis, oncology and regenerative medicine. We describe the labeling protocol with fluorescent antibodies and the electronic gating procedure in flow cytometry to score hematopoietic progenitor subsets and HSCs distribution in individual mice (Fig. 1). In addition, we describe a method to extensively enrich hematopoietic progenitors as well as long-term (LT) and short term (ST) reconstituting HSCs from pooled bone marrow cell suspensions by magnetic enrichment of cells expressing c-Kit. The resulting cell preparation can be used to sort selected subsets for in vitro and in vivo functional studies (Fig. 2).
Both trabecular osteoblasts2,3 and sinusoidal endothelium4 constitute functional niches supporting HSCs in the bone marrow. Several mechanisms in the osteoblastic niche, including a subset of N-cadherin+ osteoblasts3 and interaction of the receptor tyrosine kinase Tie2 expressed in HSCs with its ligand angiopoietin-15 concur in determining HSCs quiescence. "Hibernation" in the bone marrow is crucial to protect HSCs from replication and eventual exhaustion upon excessive cycling activity6. Exogenous stimuli acting on cells of the innate immune system such as Toll-like receptor ligands7 and interferon-&alpha;6 can also induce proliferation and differentiation of HSCs into lineage committed progenitors. Recently, a population of dormant mouse HSCs within the lin- c-Kit+ Sca-1+ CD150+ CD48- CD34- population has been described8. Sorting of cells based on CD34 expression from the hematopoietic progenitors-enriched cell suspension as described here allows the isolation of both quiescent self-renewing LT-HSCs and ST-HSCs9. A similar procedure based on depletion of lineage positive cells and sorting of LT-HSC with CD48 and Flk2 antibodies has been previously described10. In the present report we provide a protocol for the phenotypic characterization and ex vivo cell cycle analysis of hematopoietic progenitors, which can be useful for monitoring hematopoiesis in different physiological and pathological conditions. Moreover, we describe a FACS sorting procedure for HSCs, which can be used to define factors and mechanisms regulating their self-renewal, expansion and differentiation in cell biology and signal transduction assays as well as for transplantation.

A method to analyse the distribution of bone marrow hematopoietic progenitors in flow cytometry as well as to efficiently isolate highly purified hematopoietic stem cells (HSCs) is described. The isolation procedure is essentially based on magnetic enrichment of c-Kit+ cells and cell sorting to purify HSCs for cellular and molecular studies.

L&#39;analisi fenotipica e l&#39;isolamento di murini cellule staminali ematopoietiche e Lineage-committed progenitori

The bone marrow is the principal site where HSCs and more mature blood cells lineage progenitors reside and differentiate in an adult organism. HSCs ...

Lentiviral CRISPR/Cas9-Mediated Genome Editing for the Study of Hematopoietic Cells in Disease Models

Manipulating genes in hematopoietic stem cells using conventional transgenesis approaches can be time-consuming, expensive, and challenging. Benefiting from advances in genome editing technology and lentivirus-mediated transgene delivery systems, an efficient and economical method is described here that establishes mice in which genes are manipulated specifically in hematopoietic stem cells. Lentiviruses are used to transduce Cas9-expressing lineage-negative bone marrow cells with a guide RNA (gRNA) targeting specific genes and a red fluorescence reporter gene (RFP), then these cells are transplanted into lethally-irradiated C57BL/6 mice. Mice transplanted with lentivirus expressing non-targeting gRNA are used as controls. Engraftment of transduced hematopoietic stem cells are evaluated by flow cytometric analysis of RFP-positive leukocytes of peripheral blood. Using this method, ~90% transduction of myeloid cells and ~70% of lymphoid cells at 4 weeks after transplantation can be achieved. Genomic DNA is isolated from RFP-positive blood cells, and portions of the targeted site DNA are amplified by PCR to validate the genome editing. This protocol provides a high-throughput evaluation of hematopoiesis-regulatory genes and can be extended to a variety of mouse disease models with hematopoietic cell involvement.

Described are protocols for the highly efficient genome editing of murine hematopoietic stem and progenitor cells (HSPC) by the CRISPR/Cas9 system to rapidly develop mouse model systems with hematopoietic system-specific gene modifications.

Modifica del genoma mediato da Lentiviral/Cas9 per lo studio delle cellule ematopoietiche nei modelli di malattia

Manipulating genes in hematopoietic stem cells using conventional transgenesis approaches can be time-consuming, expensive, and challenging. Benefiting ...

Economical and Efficient Protocol for Isolating and Culturing Bone Marrow-derived Dendritic Cells from Mice

The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method for isolating DCs primarily involves inducing bone marrow (BM) differentiation into DCs by injecting large doses (&#62;10 ng/mL) of granulocyte-macrophage colony-stimulating factor/interleukin-4 (GM-CSF/IL-4), making the procedure complex and expensive. In this protocol, using all BM cells cultured in 10 ng/mL GM-CSF/IL-4 medium, after 3-4 half-culture exchanges, up to 2.7 x 107 CD11c+ cells&#160;(DCs) per mouse (two femurs) were harvested with a purity of 80%-95%. After 10 days in culture, the expression of CD11c, CD80, and MHC II increased, whereas the number of cells decreased. The number of cells peaked after 7 days of culture. Moreover, this method only took 10 min to harvest all bone marrow cells, and a high number of DCs were obtained after 1 week of culture.

Here, we present an economical and efficient method to isolate and generate high-purity bone marrow-derived dendritic cells from mice after 7 days of culture with 10 ng/mL GM-CSF/IL-4.

Protocollo economico ed efficiente per l'isolamento e la coltura di cellule dendritiche derivate dal midollo osseo da topi

The demand for dendritic cells (DCs) is gradually increasing as immunology research advances. However, DCs are rare in all tissues. The traditional method ...

Ottimizzazione dell'isolamento delle cellule stromali dal timo e dal midollo osseo

Stromal Cell Isolation From Hematopoietic Organs

Single-cell sequencing has enabled the mapping of heterogeneous cell populations in the stroma of hematopoietic organs. These methodologies provide a lens through which to study previously unresolved heterogeneity at steady state, as well as changes in cell type representation induced by extrinsic stresses or during aging. Here, we present step-wise protocols for the isolation of high-quality stromal cell populations from murine and human thymus, as well as murine bone and bone marrow. Cells isolated through these protocols are suitable for generating high-quality single-cell multiomics datasets. The impacts of sample digestion, hematopoietic lineage depletion, FACS analysis/sorting, and how these factors influence compatibility with single-cell sequencing are discussed here. With examples of FACS profiles indicating successful and inefficient dissociation and downstream stromal cell yields in post-sequencing analysis, recognizable pointers for users are provided. Considering the specific requirements of stromal cells is crucial for acquiring high-quality and reproducible results that can advance knowledge in the field.

Autore in primo piano: Avanzamento della ricerca ematopoietica utilizzando l'isolamento delle cellule stromali per il sequenziamento di singole cellule

Here we present protocols that enable isolation of stromal cells from murine bone, bone marrow, thymus and human thymic tissue compatible with single-cell multiomics.

Isolamento delle cellule stromali da organi ematopoietici

Single-cell sequencing has enabled the mapping of heterogeneous cell populations in the stroma of hematopoietic organs. These methodologies provide a lens ...

Expression of Fluorescent Fusion Proteins in Murine Bone Marrow-derived Dendritic Cells and Macrophages

Dendritic cells and macrophages are crucial cells that form the first line of defense against pathogens. They also play important roles in the initiation of an adaptive immune response. Experimental work with these cells is rather challenging. Their abundance in organs and tissues is relatively low. As a result, they cannot be isolated in large numbers. They are also difficult to transfect with cDNA constructs. In the murine model, these problems can be partially overcome by in vitro differentiation from bone marrow progenitors in the presence of M-CSF for macrophages or GM-CSF for dendritic cells. In this way, it is possible to obtain large amounts of these cells from very few animals. Moreover, bone marrow progenitors can be transduced with retroviral vectors carrying cDNA constructs during early stages of cultivation prior to their differentiation into bone marrow derived dendritic cells and macrophages. Thus, retroviral transduction followed by differentiation in vitro can be used to express various cDNA constructs in these cells. The ability to express ectopic proteins substantially extends the range of experiments that can be performed on these cells, including live cell imaging of fluorescent proteins, tandem purifications for interactome analyses, structure-function analyses, monitoring of cellular functions with biosensors and many others. In this article, we describe a detailed protocol for retroviral transduction of murine bone marrow derived dendritic cells and macrophages with vectors coding for fluorescently-tagged proteins. On the example of two adaptor proteins, OPAL1 and PSTPIP2, we demonstrate its practical application in flow cytometry and microscopy. We also discuss the advantages and limitations of this approach.

In this article, we provide a detailed protocol for the expression of fluorescent fusion proteins in murine bone marrow derived dendritic cells and macrophages. The method is based on the transduction of bone marrow progenitors with retroviral constructs followed by differentiation into macrophages and dendritic cells in vitro.

Espressione di proteine di fusione fluorescenti in macrofagi e cellule dendritiche derivate da midollo osseo Murine

Dendritic cells and macrophages are crucial cells that form the first line of defense against pathogens. They also play important roles in the initiation ...

La preparazione di colture primarie di cellule emopoietiche dal midollo osseo murino per elettroporazione

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Esplora Altri Video

Capitoli in questo video

Rottura del Gene altamente efficiente di cellule progenitrici ematopoietiche Murine ed umane di CRISPR/Cas9

Isolamento e iniezione endovenosa di Murine derivate dal midollo osseo monociti

Utilizzando il sistema di elettroporazione Gene Pulser MXcell per trasfettare cellule primarie con alta efficienza

L'analisi fenotipica e l'isolamento di murini cellule staminali ematopoietiche e Lineage-committed progenitori

Modifica del genoma mediato da Lentiviral/Cas9 per lo studio delle cellule ematopoietiche nei modelli di malattia

Protocollo economico ed efficiente per l'isolamento e la coltura di cellule dendritiche derivate dal midollo osseo da topi

Isolamento delle cellule stromali da organi ematopoietici

Isolamento delle cellule stromali da organi ematopoietici

Isolamento delle cellule stromali da organi ematopoietici

Espressione di proteine di fusione fluorescenti in macrofagi e cellule dendritiche derivate da midollo osseo Murine