Journal of Cell Science & Therapy
Open Access
ISSN: 2157-7013
+44 1300 500008
ISSN: 2157-7013
+44 1300 500008
De Schauwer Catharina, Evelyne Meyer, Gerlinde R. Van de Walle, Guy Wouters and Ann Van Soom
Accepted Abstracts: J Cell Sci Ther
Th e therapeutic potential of mesenchymal stromal cells (MSC) for cellular therapy has generated an increasing interest in this type of research. In human, as well as in veterinary medicine, considerable research has been performed on the cryopreservation of expanded MSC, but little information is available on the possibility to cryopreserve the original mononuclear cell fraction. Th e present study describes a protocol to successfully expand equine MSC aft er cryopreserving the mononuclear cell fraction of umbilical cord blood (UCB). To this end, the mononuclear cell fraction was isolated from 5 UCB samples using a Percoll gradient and cryopreserved in standard 1.8ml cryotubes at a concentration of 1-2×10 6 cells per ml cold freezing solution. Cells were kept frozen for a minimum of three weeks before thawing. Frozen cryotubes were thawed au bain marie at 37°C. Cell viability aft er thawing varied around 98%. Approximately 4 × 10 6 cells were seeded in a T25 culture fl ask in culture medium containing low-glucose DMEM, 30% FCS, 10 - 7 M low dexamethazone, 50 μg/mL gentamycine, 10 μl/ ml antibiotic antimycotic solution, 250 ng/mL fungizone and 2 mM ultraglutamine. In 4 out of 5 samples, adherent spindle-shaped cell colonies occurred within 9.8 ± 3.2 days and 80% confl uency was reached aft er 16.3 ± 2.2 days of incubation at 38.5°C and 5% CO 2 . Aft er three passages, undiff erentiated MSC were immunophenotyped using multi-color fl ow cytometry. In conclusion, equine MSC can be cultured successfully aft er cryopreservation of the isolated mononuclear cell fraction, an approach that is time- as well as cost-effi cient