Evaluation of bone marrow mesenchymal stem cell standard cryopreservation procedure efficiency
K A T Carvalho1, C C Cury, L Oliveira
1Cellular Culture Experimental Laboratory, PUCPR, Brazil. katherinecarv@gmail.com
Transplantation Proceedings
|May 6, 2008
Summary
Standard cryopreservation methods are inefficient for adult bone marrow mesenchymal stem cells. Flow cytometry is more sensitive than the trypan blue test for assessing cell viability after cryopreservation.
Area of Science:
- Stem Cell Biology
- Cryobiology
- Regenerative Medicine
Background:
- Mesenchymal stem cells (MSCs) from bone marrow hold significant potential for tissue regeneration and clinical applications.
- Cryopreservation allows for long-term storage of various cell types, including MSCs.
Purpose of the Study:
- To evaluate the efficacy of standard cryopreservation techniques for adult bone marrow-derived MSCs.
- To compare the sensitivity of different viability assays post-cryopreservation.
Main Methods:
- Adult rat bone marrow mononuclear cells were cultured to obtain MSCs.
- Cells were cryopreserved in liquid nitrogen vapor phase for one month.
- Viability was assessed using trypan blue exclusion and flow cytometry (annexin V/7-AAD) before and after thawing.
Main Results:
- Trypan blue exclusion showed high viability (94.76% pre, 90.58% post-thaw).
- Flow cytometry indicated a significant decrease in viability (85.52% pre, 66.25% post-thaw).
- Flow cytometry demonstrated greater sensitivity in detecting cell death.
Conclusions:
- Standard cryopreservation protocols are inadequate for preserving the viability of bone marrow MSCs.
- Flow cytometry is a more sensitive method for assessing MSC viability after cryopreservation compared to trypan blue exclusion.
