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Thawing and expansion of frozen cells, each aliquot

CPT4 code

Name of the Procedure:

Thawing and Expansion of Frozen Cells (Cryopreserved Cell Recovery)


Thawing and expansion of frozen cells is a lab procedure where frozen cell samples, preserved at very low temperatures, are carefully thawed and cultured to increase their numbers. This process is typically conducted to prepare cells for further research, medical treatments, or diagnostic purposes.


The procedure is used to revive and expand frozen cells for various purposes, including research, therapeutic treatments, and diagnostic tests. The goal is to obtain a sufficient quantity of viable cells while maintaining their original characteristics and function.


  • Research requiring specific cell types previously cryopreserved.
  • Medical treatments, such as stem cell therapies.
  • Diagnostic applications needing large quantities of specific cell types.


  • Ensure a sterile working environment to avoid contamination.
  • Pre-warm all required reagents and culture media to the appropriate temperatures.
  • Confirm all equipment, such as water baths and incubators, is functioning correctly.

Procedure Description

  1. Retrieve a frozen cell aliquot from cryostorage.
  2. Quickly transfer the aliquot to a 37°C water bath for rapid thawing, agitating gently until just thawed.
  3. Immediately transfer the contents to a sterile tube containing pre-warmed culture medium.
  4. Centrifuge the cells to pellet them, remove the supernatant, and resuspend the cells in fresh medium.
  5. Plate the cells into appropriate culture vessels and place them in an incubator set to optimal conditions for the specific cell type.
  6. Monitor the cells for growth and viability, changing the medium as needed.

Tools/Equipment Used:

  • 37°C water bath
  • Sterile tubes and pipettes
  • Centrifuge
  • Incubators
  • Culture media and vessels

Anesthesia or Sedation:

  • Not applicable


The thawing process takes approximately 15-30 minutes, but the expansion phase where cells multiply can take several days to weeks, depending on the cell type.


This procedure is typically performed in a laboratory setting, such as a research lab or a clinical cell processing facility.


  • Laboratory technologists or scientists
  • Researchers
  • Laboratory assistants

Risks and Complications

  • Contamination of cell cultures
  • Reduction in cell viability if thawing process is too slow or not conducted properly
  • Potential cellular changes or loss of function during the thawing or expansion process


  • Enables the use of previously stored cell samples for research or therapeutic purposes.
  • Provides a viable quantity of cells necessary for advanced studies or treatments.
  • Immediate availability of specific cell types as needed.


  • Post-thaw care involves monitoring cell cultures for contamination and ensuring optimal growth conditions.
  • Fresh medium changes and occasional splitting of cells may be required.
  • Full recovery and expansion can take from a few days to several weeks.


  • Starting from freshly isolated cells, which can be time-consuming and may not always be possible.
  • Utilizing pre-expanded cell lines, although they may not be identical to the cryopreserved samples.

Pros of Alternatives:

  • Fresh cells may have better viability.
  • Pre-expanded cell lines are immediately available.

Cons of Alternatives:

  • Fresh isolation can be impractical and risk patient discomfort.
  • Pre-expanded lines might not meet the specific experimental needs.

Patient Experience

Since this is a laboratory procedure with no direct patient involvement, patients might not experience any direct effects. However, it's essential for patients receiving treatments involving expanded cells (such as stem cell therapy) to understand the procedure was meticulously handled to ensure cell viability and effectiveness.

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