Human Adipose-Derived Mesenchymal Stem Cells Cryopreservation and Thawing Decrease α4-Integrin Expression
Ana Carolina Irioda1, Rafael Cassilha2, Larissa Zocche3
1Cell Therapy and Biotechnology in Regenerative Medicine Research Group, Pelé Pequeno Príncipe Institute, Avenida Silva Jardim 1632, 80250-200 Curitiba, PR, Brazil; Bioprocess Engineering and Biotechnology Department, Federal University of Paraná, Avenida Coronel Francisco Heráclito dos Santos 210, 81531-970 Curitiba, PR, Brazil.
Abstract:
Aim. The effects of cryopreservation on adipose tissue-derived mesenchymal stem cells are not clearly documented, as there is a growing body of evidence about the importance of adipose-derived mesenchymal stem cells for regenerative therapies. The aim of this study was to analyze human adipose tissue-derived mesenchymal stem cells phenotypic expression (CD34, CD45, CD73, CD90, CD105, and CD49d), colony forming unit ability, viability, and differentiation potential before and after cryopreservation. Materials and Methods. 12 samples of the adipose tissue were collected from a healthy donor using the liposuction technique. The cell isolation was performed by enzymatic digestion and then the cells were cultured up to passage 2. Before and after cryopreservation the immunophenotype, cellular viability analysis by flow cytometer, colony forming units ability, differentiation potential into adipocytes and osteoblasts as demonstrated by Oil Red O and Alizarin Red staining, respectively. Results. The immunophenotypic markers expression was largely preserved, and their multipotency was maintained. However, after cryopreservation, the cells decreased α4-integrin expression (CD49d), cell viability, and number of colony forming units. Conclusions. These findings suggest that ADMSC transplanted after cryopreservation might compromise the retention of transplanted cells in the host tissue. Therefore, further studies are warranted to standardize protocols related to cryopreservation to attain full benefits of stem cell therapy.
Insights
Cryopreservation impacts adipose tissue-derived mesenchymal stem cells (ADMSCs), reducing viability and colony formation. Preserving ADMSCs requires optimized protocols to maintain their therapeutic potential for regenerative medicine.
Area of Science:
- Regenerative Medicine
- Stem Cell Biology
- Tissue Engineering
Background:
- Adipose tissue-derived mesenchymal stem cells (ADMSCs) are crucial for regenerative therapies.
- Cryopreservation effects on ADMSC function require thorough investigation.
Purpose of the Study:
- To evaluate the impact of cryopreservation on ADMSC phenotype, viability, and differentiation.
- To assess changes in immunophenotypic markers (CD34, CD45, CD73, CD90, CD105, CD49d) post-cryopreservation.
Main Methods:
- Human ADMSCs isolated from adipose tissue via liposuction.
- Analysis of immunophenotype, viability, colony-forming units, and differentiation potential before and after cryopreservation.
Main Results:
- Immunophenotypic marker expression was largely maintained.
- Cryopreservation led to decreased CD49d expression, reduced cell viability, and fewer colony-forming units.
Conclusions:
- Cryopreserved ADMSCs may have compromised retention in host tissues.
- Further research is needed to optimize cryopreservation protocols for ADMSCs in regenerative medicine.


