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CrossNetwork PBMC Processing Standard Operating Procedure Appendix A: HVTN PBMC Processing Worksheet Specimen Processing Laboratory: Participant ID (PTID): Visit: Protocol: Collection Date: Time:.

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To obtain T cells for CAR-T cell production, blood is collected and leukapheresis is performed to isolate and extract peripheral mononuclear blood cells (PBMCs, e.g., leukocytes and lymphocytes). The T cells in this PMBC fraction are then separated out using cell processors or centrifugation-based cell separators.

The PBMC isolation protocol involves directly separating lymphocytes from whole blood. Whole blood is the blood that flows through the human body — the raw fluid with no components removed or separated. This is composed primarily of plasma, platelets, red blood cells, and PBMCs.

This chapter deals with T lymphocytes derived from peripheral blood mononuclear cells (PBMCs). T cells comprise, approximately, 45–70 % of PBMCs in human peripheral blood, with a count in healthy subjects in the range of 1 million cells/mL.

The most common PBMC isolation method involves using a density gradient medium (e.g. Ficoll™ or Lymphoprep™) and centrifugation. This method takes advantage of the differences in density between the cells in blood and the density gradient medium.

PBMCs include lymphocytes (T cells, B cells, and NK cells), monocytes, and dendritic cells.

Peripheral blood mononuclear cell (PBMC) isolation is commonly done via density gradient centrifugation over Ficoll-, a labor-intensive procedure that requires skilled technicians and can contribute to sample variability.

The frequencies of cell types within the lymphocyte population include 70 – 85% CD3+ T cells (45 – 70% of PBMC), 5 – 20% B cells (up to 15% of PBMC), and 5 – 20% NK cells (up to 15% of PBMC). The CD3+ compartment is composed of CD4 (25 – 60% of PBMC) and CD8 T cells (5 – 30% of PBMC), in a roughly 2:1 ratio.

Because erythrocytes and granulocytes are much more fragile to freezing and thawing, PBMC isolation is a common prerequisite to cryopreservation of blood cells; and the most common method for PBMC isolation is density gradient centrifugation using Ficoll-, a high molecular weight carbohydrate solution.

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Form Packages
Adoption
Bankruptcy
Contractors
Divorce
Home Sales
Employment
Identity Theft
Incorporation
Landlord Tenant
Living Trust
Name Change
Personal Planning
Small Business
Wills & Estates
Packages A-Z
Form Categories
Affidavits
Bankruptcy
Bill of Sale
Corporate - LLC
Divorce
Employment
Identity Theft
Internet Technology
Landlord Tenant
Living Wills
Name Change
Power of Attorney
Real Estate
Small Estates
Wills
All Forms
Forms A-Z
Form Library
Customer Service
Terms of Service
DMCA Policy
About Us
Blog
Affiliates
Contact Us
Privacy Notice
Delete My Account
Site Map
All Forms
Search all Forms
Industries
Forms in Spanish
Localized Forms
Legal Guides
Real Estate Handbook
All Guides
Prepared for You
Notarize
Incorporation services
Our Customers
For Consumers
For Small Business
For Attorneys
Our Sites
US Legal Forms
USLegal
FormsPass
pdfFiller
signNow
airSlate workflows
DocHub
Instapage
Social Media
Call us now toll free:
1-877-389-0141
As seen in:
  • USA Today logo picture
  • CBC News logo picture
  • LA Times logo picture
  • The Washington Post logo picture
  • AP logo picture
  • Forbes logo picture
© Copyright 1997-2025
airSlate Legal Forms, Inc.
3720 Flowood Dr, Flowood, Mississippi 39232