Name: homing of hematopoietic cells to the bone marrow
Uploaded: 2009-03-18T00:00:00
Description: Watch this Scientific Journal Video about Homing of Hematopoietic Cells to the Bone Marrow at JoVE.com

<p class="jove_content">This protocol was tested and optimized in the laboratory of Dr. David Scadden. Funding is provided by the Dana Farber Cancer Institute/Massachusetts General Hospital Clinical Hematology and Oncology Fellowship Program.</p>

<ol>
<li> We begin the homing experiment by extracting cells to be used in the experiment. For this current experiment we are interested in finding out if the homing of whole bone marrow to the niches in the long bones of recipient animals is different for bone marrow cells extracted from WT animals such as C57BL/6J (WT) and those that are transgenic knockouts for Lysophosphatidic Receptor 1 (LPAR1) (KO).</li>
<li>Before we start the experiment,  we will collect the materials we need for this experiment.  In our lab all mouse dissections are ca...

<ol>
	<li>Adams, G. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cell+engraftment+at+the+endosteal+niche+is+specified+by+the+calcium-sensing+receptor.">Stem cell engraftment at the endosteal niche is specified by the calcium-sensing receptor.</a> <em style='font-style: italic'>Nature</em>. <b>439</b> (7076), 599-603 (2006).</li><li>Broxmeyer, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemokines+in+hematopoiesis.">Chemokines in hematopoiesis.</a> <em style='font-style: italic'>Curr Opin Hematol</em>. <b>15</b> (1), 49-58 (2008).</li></ol>

<p class="jove_content">Homing is the process whereby bone marrow cells including stem cells, progenitor cells and differentiated cells, their way  into the bone marrow after being injected into the blood stream or bone marrow cavity of mice. As is obvious from the definition, a scientist may chose to study the homing of hematopoietic stem cells, progenitor cells or differentiated cells  identified by immunophenotyping and isolated by cell sorting. Typically scientists inject bone marrow cells depleted for lineage markers and hence enriched for...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>DPBS (Dulbecco&#x2019;s Phosphate Buffered Saline</td>
<td>&nbsp;</td>
<td>Mediatech, Inc</td>
<td>21-031-CV</td>
<td>&nbsp;</td>
<tr>
<td>Sterile Fetal Bovine Serum</td>
<td>&nbsp;</td>
<td>Valley Biomedical, Inc</td>
<td>BS3033</td>
<td>&nbsp;</td>
<tr>
<td>0.5M EDTA (pH 8.0)</td>
<td>&nbsp;</td>
<td>Boston BioProducts</td>
<td>BM-150</td>
<td>&nbsp;</td>
<tr>
<td>Tryphan Blue solution</td>
<td>&nbsp;</td>
<td>Mediatech, Inc</td>
<td>25-900-CI</td>
<td>&nbsp;</td>
<tr>
<td>ACK Lysing Buffer</td>
<td>&nbsp;</td>
<td>Lonza</td>
<td>10-548E</td>
<td>&nbsp;</td>
<tr>
<td>Isopropranolol U.S.P.</td>
<td>&nbsp;</td>
<td>Denison Pharmaceuticals, Inc</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Vybrant DiD cell-labelling solution</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>V22887</td>
<td>&nbsp;</td>
<tr>
<td>Vybrant DiI cell-labelling solution</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>V22885</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

homing of hematopoietic cells to the bone marrow

<p class="jove_content">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]</p>
<p class="jove_content">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.</p>

Watch this Scientific Journal Video about Homing of Hematopoietic Cells to the Bone Marrow at JoVE.com

Homing of Hematopoietic Cells to the Bone Marrow

<p class="jove_content">This article describes a protocol used to study the homing of hematopoietic cells to their niches in the bone marrow.</p>

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

Research

JoVE Journal

Biology

Isolation and Analysis of Hematopoietic Stem Cells from the Placenta

Hematopoietic stem cells (HSCs) have the ability to self-renew and generate all cell types of the blood lineages throughout the lifetime of an individual. ...

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 ...

Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging

Several methods for the preparation of murine dendritic cells can be found in the literature. Here, we present a method that produces greater than 85% ...

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

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

Lentiviral-mediated Knockdown During <em>Ex Vivo</em> Erythropoiesis of Human Hematopoietic Stem Cells

Lentiviral-mediated Knockdown During Ex Vivo Erythropoiesis of Human Hematopoietic Stem Cells

Erythropoiesis is a commonly used model system to study cell differentiation. During erythropoiesis, pluripotent adult human hematopoietic stem cells ...

<em>In Vivo</em> 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

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 ...

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 ...

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 ...

Proplatelet Formation Dynamics of Mouse Fresh Bone Marrow Explants

The last stage of megakaryopoiesis leads to cytoplasmic extensions from mature megakaryocytes, the so-called proplatelets. Much has been learned about the ...