Resources
Cross-Network PBMC Processing SOP

Procedures for the Isolation and Cryopreservation of PBMC

The Cross-Network Peripheral Blood Mononuclear Cell (PBMC) Processing Standard Operating Procedure (SOP) provides instructions for processing PBMC at network (ACTG, HPTN, HVTN, IMPAACT, and MTN) site-affiliated laboratories, and is intended to simplify and clarify PBMC processing, especially at shared sites. Use of the Cross-Network PBMC Processing SOP is required for all laboratories cryopreserving PBMC for ACTG, IMPAACT, HPTN, HVTN and MTN.

Current Versions

Before using the SOP for the first time, thoroughly review all sections that apply to your network(s) for important notes and details. When you are familiar with the procedures, you may use the quick guides found in the appendices of the Cross-Network PBMC Processing SOP.

English

Cross-Network PBMC Processing SOP v6.0
Version 6.0 is effective as of April 26, 2018.

Spanish (South American)

Procesamiento de PBMC entre redes SOP v6.0 (Español)
Version 6.0 is effective as of April 30, 2018.

Portuguese (Brazilian)

Processamento de PBMC interrede SOP v6.0 (Português)
Version 6.0 is effective as of April 30, 2018.

Thai

Version Change Summary

Version 6.0 differs from 5.2 by the following changes: 

  • Section 8.1.4: Additional requirements captured in October 7, 2016 memo for ACTG and IMPAACT labs​ only added. Additional cryovial-related requirements for all networks also added.
  • Section 18.2.1: Additional guidelines related to freezer set point temperatures added.
  • Appendix G: Added additional information to the Cryovial section related to use of the SARSTEDT Screw cap micro tube.

Please refer to Appendix H of the SOP for a complete history of changes.

SOP Modification

Not all sections of the Cross-Network PBMC Processing SOP are needed in my laboratory. May we modify it?

The Laboratory Director is responsible for the implementation of this HANC SOP or laboratory-specific SOP and for ensuring that all appropriate personnel are trained. A laboratory-specific SOP must:

  • Include, without procedural modification, the portions of the current version of the Cross-Network PBMC Processing SOP that are used within the network site-affiliated laboratory
  • Reference the current version of the Cross-Network PBMC Processing SOP

Note: For laboratories processing HVTN PBMCs, the laboratory must use the HANC PBMC SOP as written or the HVTN-specific version of the HANC SOP.

To develop a laboratory-specific SOP, click on the appropriate version and it will open as a new document. Save it with the desired name and make the necessary changes.

Cross-Network PBMC Processing SOP v6.0 Template (dotx)
Procesamiento de PBMC entre redes SOP v6.0​ Plantilla (dotx)
Processamento de PBMC interrede SOP v6.0 Modelo (dotx)
กระบวนการระหว่างเครือข่าย PBMC SOP v6.0 แบบv (dotx)

Results Reporting

The requirements for reporting results have changed with version 3.0. Use of a PBMC Processing Worksheet and the Laboratory Data Management System (LDMS) is now required for all networks to track the timing of processing, calculations and documentation of problems that arise during processing. Be sure to review section 5 Reporting Results for the new requirements for use of a PBMC Processing Worksheet and the LDMS.

HVTN

The use of the HVTN PBMC Processing Worksheet in its entirety is required. If a protocol-specific PBMC worksheet is not required in the “HVTN Protocol Specific Processing Instructions,” the generic worksheet below and in Appendix A of the SOP may be used.

HVTN PBMC Processing Worksheet v5.1
Hoja de trabajo para procesamiento de PBMC v5.1

ACTG, HPTN, IMPAACT, MTN

The laboratory may use the HVTN PBMC Processing Worksheet, or modify it as appropriate for network policy and laboratory’s procedures. If the lab chooses to develop its own PBMC Processing Worksheet and supplementary tracking materials (such as the LDMS, or a separate worksheet or log) the laboratory will use the guidelines in section 5 Reporting Results of the SOP.

Several PBMC Processing Worksheet templates that meet the requirements are provided below. To develop a laboratory-specific worksheet, click on the appropriate version and it will open as a new document. Save it with the desired name and make the necessary changes.

PBMC Processing Worksheet Template (dotx; similar to HVTN PBMC Processing Worksheet v5.1)
Hoja de trabajo para procesamiento de PBMC Plantilla v5.1 (dotx)
Planilha de processamento de CMSP Modelo v5.1 (dotx)
ตารางการเตรียม PBMC แบบv 5.1 (dotx)
PBMC Processing Worksheet Template 2 (rtf)
PBMC Processing Worksheet Template 3 (rtf)

Example Manual Counting SOP

Below is the Example PBMC Hemacytometer Counting SOP. Note that this is an example - not a required - SOP. It must be tailored to laboratory-specific and/or study-specific requirements before it can be used.

To develop a laboratory-specific PBMC Hemacytometer Counting SOP, save (don't open) the file to your computer. Open the document, save it as a new file, and make the necessary changes.

Example PBMC Hemacytometer Counting SOP (rtf)