Abstract
Mesenchymal stem cells (MSCs) are pluripotential cells present in marrow, which have the ability to differentiate into osteoblasts, chrondrocytes and adipocytes. Potential skeletal tissue engineering uses include healing bone defects, spinal fusion and revision arthoplasty surgery. A means of storing viable mesenchymal stem cells is necessary in order for these cells to be readily available for clinical use. The aim of this study was to determine whether cryopreservation has any effect on the osteogenic potential of human bone marrow derived MSCs.
Five normal iliac crest bone marrow aspirates were obtained following informed consent from patients. Each aspirate was divided into two equal samples. Ficoll-separation was used to isolate the MSCs. The fresh MSCs from one sample were cryopreserved, while the other was cultured as a control population. To assess the osteogenic potential of the MSCs after cryopreservation a sample of cells from each population was cultured with osteogenic supplements and the increase in alkaline phosphatase (ALP) and osteocalcin production was compared.
Cryopreservation was not observed to effect the primary cultures of MSCs, which became confluent after a similar period in culture (12–14 days), forming colonies with recognized MSCs morphology. The expression of ALP and osteocalcin after stimulating the MSCs to differentiate with osteogenic supplements, was not significantly altered by the cryopreservation process (P> 0.05).
In conclusion MSCs obtained from fresh human bone marrow aspirates can be cryopreserved without compromise to their proliferation rate or osteogenic potential, confirming that this is a useful means of storing viable cells for future clinical use.
Abstracts prepared by Dr P E Watkins, Hodgkin Building, Guys Campus, King’s College London.
