A Reliable Method for Thawing Primary AML and CMML Mononuclear Cells to Preserve Viability and Function

Area of Science:

• Hematology
• Cell Biology
• Cryobiology

Background:

• Human mononuclear cells from blood and bone marrow are crucial for studying hematological malignancies like AML and CMML.
• Cryopreservation allows long-term storage of these valuable patient samples.
• Existing thawing protocols often struggle with poor post-thaw viability and recovery, especially for sensitive blast and monocyte populations.

Purpose of the Study:

• To develop and validate a reliable protocol for thawing cryopreserved acute myeloid leukemia (AML) and chronic myelomonocytic leukemia (CMML) mononuclear cells.
• To optimize post-thaw viability, cell recovery, and functional integrity of these hematopoietic cells.
• To provide a robust method applicable to fragile or monocyte-rich patient-derived samples.

Main Methods:

• Controlled dilution of cryopreserved cells using anticoagulant-supplemented thaw buffer.
• Inclusion of DNase I treatment to prevent cell aggregation.
• Gentle resuspension techniques to maintain cell integrity.

Main Results:

• Achieved consistent post-thaw viability exceeding 80%.
• Demonstrated a mean cell recovery rate of 55.6% across samples.
• Confirmed retention of functional capacity (colony-forming assays) and immunophenotypic characteristics (flow cytometry).

Conclusions:

• The described thawing protocol is robust and reproducible for cryopreserved AML and CMML mononuclear cells.
• This method effectively preserves cell viability, recovery, and function, overcoming challenges with fragile cell types.
• The protocol has broad applicability for thawing other patient-derived hematopoietic samples prone to aggregation or poor recovery.