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<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1000 &#181;L Tips</td>
			<td>Gilson</td>
			<td>F174501</td>
			<td>Approximately 3 tips needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>15 mL tube</td>
			<td>Biopioneeer</td>
			<td>CNT-15</td>
			<td>To hold Solutions 2/3 
			2 needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>2 mL Cryovials</td>
			<td>Globe Scientific</td>
			<td>3012</td>
			<td>Approximately 40 cryovials needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>20 &#181;L Tips</td>
			<td>Gilson</td>
			<td>F174201</td>
			<td>For use in cell counting; volume needed is dependent on method of cell counting used. 1 tip is needed per 1 mL of unpooled frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>50 mL Conical Tube</td>
			<td>CEM Corporation</td>
			<td>50-187-7683</td>
			<td>2 needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection 
			3 needed per 1 mL of unpooled frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>CD14 Antibody</td>
			<td>Biolegend</td>
			<td>325621</td>
			<td></td>
		</tr>
		<tr>
			<td>CD16 Antibody</td>
			<td>Biolegend</td>
			<td>360723</td>
			<td></td>
		</tr>
		<tr>
			<td>CD19 Antibody</td>
			<td>Biolegend</td>
			<td>363007</td>
			<td></td>
		</tr>
		<tr>
			<td>CD3 Antibody</td>
			<td>Biolegend</td>
			<td>300405</td>
			<td></td>
		</tr>
		<tr>
			<td>CD56 Antibody</td>
			<td>Biolegend</td>
			<td>318303</td>
			<td></td>
		</tr>
		<tr>
			<td>CD66b Antibody</td>
			<td>Biolegend</td>
			<td>305103</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell Strainer</td>
			<td>Biopioneeer</td>
			<td>DGN258367</td>
			<td>1 needed per 1 mL of unpooled frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>CPT Mononuclear Cell Preparation Tube</td>
			<td>BD Biosciences</td>
			<td>362753</td>
			<td>6 tubes needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Cryo Freezer Box</td>
			<td>Southern Labware</td>
			<td>SB2CC-81</td>
			<td>Holds 81 tubes 
			1 needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Cryotube Rack</td>
			<td>Fisherbrand</td>
			<td>05-669-45</td>
			<td>Holds up to 50 cryovials</td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Invitrogen</td>
			<td>15575020</td>
			<td>Approximately 2.5 mL needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>DNase/RNase-Free Distilled Water</td>
			<td>Invitrogen</td>
			<td>10977015</td>
			<td>Approximately 2 mL needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection 
			Approximately 9 mL needed per 1 mL of frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>EDTA Tubes</td>
			<td>BD Biosciences</td>
			<td>366643</td>
			<td>1 tube needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Flavopiridol</td>
			<td>Sigma-Aldrich</td>
			<td>F3055-5MG</td>
			<td></td>
		</tr>
		<tr>
			<td>HLA-DR Antibody</td>
			<td>Biolegend</td>
			<td>307609</td>
			<td></td>
		</tr>
		<tr>
			<td>Human Serum Albumin</td>
			<td>GeminiBio</td>
			<td>800-120</td>
			<td>Approximately 25 mL needed per patient or treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Human Trustain FcX</td>
			<td>Biolegend</td>
			<td>422301</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropanol</td>
			<td>Sigma-Aldrich</td>
			<td>W292912-1KG-K</td>
			<td>For use in Mr. Frosty</td>
		</tr>
		<tr>
			<td>Label Printer</td>
			<td>Phomemo</td>
			<td>M110-WH</td>
			<td></td>
		</tr>
		<tr>
			<td>Live-Dead Stain</td>
			<td>Biolegend</td>
			<td>423105</td>
			<td></td>
		</tr>
		<tr>
			<td>Mr. Frosty</td>
			<td>Thermo Scientific</td>
			<td>5100-0001</td>
			<td>Holds up to 18 tubes. 
			2 Mr. Frostys needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Multidispense Pipette</td>
			<td>Brandtech</td>
			<td>705110</td>
			<td></td>
		</tr>
		<tr>
			<td>Multidispense Pipette Tips</td>
			<td>Brandtech</td>
			<td>705744</td>
			<td>Approximately 3 tips needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>P1000 Pipette</td>
			<td>Gilson</td>
			<td>F144059M</td>
			<td></td>
		</tr>
		<tr>
			<td>P20 Pipette</td>
			<td>Gilson</td>
			<td>F144056M</td>
			<td>For use in cell counting; volume needed is dependent on method of cell counting used.</td>
		</tr>
		<tr>
			<td>Printer Labels</td>
			<td>Phomemo</td>
			<td>PM-M110-3020</td>
			<td>Approximately 45 labels needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection 
			Approximately 5 labels needed per 1 mL of unpooled frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>RNase-Free EDTA (0.5 M)</td>
			<td>Invitrogen</td>
			<td>AM9260G</td>
			<td>Approximately 40 &#181;L needed per 1 mL of frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>RNase-Free PBS (10X)</td>
			<td>Invitrogen</td>
			<td>AM9625</td>
			<td>Approximately 1 mL needed per 1 mL of frozen PBMCs to be defrosted</td>
		</tr>
		<tr>
			<td>RNasin</td>
			<td>Promega</td>
			<td>N2111</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI</td>
			<td>Corning</td>
			<td>10-040-CV</td>
			<td>Approximately 80 mL needed per patient or treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Transfer Pipettes</td>
			<td>Fisherbrand</td>
			<td>13-711-7M</td>
			<td>Approximately 5 needed per patient/treatment condition for PBMC/Plasma/Buffy Coat Collection</td>
		</tr>
		<tr>
			<td>Tube Holder</td>
			<td>Endicott-Seymour</td>
			<td>14-781-15</td>
			<td>Holds up to 80 CPT/EDTA Tubes</td>
		</tr>
	</tbody>
</table>

isolation and cryopreservation of highly viable human peripheral blood mononuclear cells from whole blood: a guide for beginners

Isolering och kryokonservering av mycket livskraftiga humana mononukleära celler i perifert blod från helblod: En guide för nybörjare

Peripheral blood mononuclear cells (PBMCs) are commonly used in biomedical research on the immune system and its response to disease and pathogens. This ...

isolation-cryopreservation-highly-viable-human-peripheral-blood

Research

JoVE Journal

Medicine

Isolation and Cryopreservation of Highly Viable Human Peripheral Blood Mononuclear Cells From Whole Blood: A Guide for Beginners

1.5K Views.  University of California San Diego. In this protocol, we aim to provide an easy-to-follow, step-by-step tutorial on how to collect and store high viability peripheral blood and mononuclear cells, or PBMCs, from whole blood. PBMCs include lymphocytes, dendritic cells, and monocytes. Bulk PBMCs can be processed to isolate individual cell types and are commonly used in a wide variety of experiments and assays, including RNA sequencing and flow cytometry.In addition to PBMCs, we also demonstrate how to collect EDTA plasma an...