Comparative studies of mesenchymal stem cells derived from different cord tissue compartments - The influence of cryopreservation and growth media.
BACKGROUND
We have identified some critical aspects concerning umbilical cord tissue mesenchymal stem cells: the lack of standards for cell isolation, expansion and cryopreservation, the lack of unanimous opinions upon their multilineage differentiation potential and the existence of very few results related to the functional characterization of the cells isolated from cryopreserved umbilical cord tissue. Umbilical cord tissue cryopreservation appears to be the optimal solution for umbilical cord tissue mesenchymal stem cells storage for future clinical use. Umbilical cord tissue cryopreservation allows mesenchymal stem cells isolation before expected use, according with the specific clinical applications, by different customized isolation and expansion protocols agreed by cell therapy institutions.


METHODS
Using an optimized protocol for umbilical cord tissue cryopreservation in autologous cord blood plasma, isolation explant method and growth media supplemented with FBS or human serum, we performed comparative studies with respect to the characteristics of mesenchymal stem cells (MSC) isolated from different compartments of the same umbilical cord tissue such as Wharton's jelly, vein, arteries, before cryopreservation (pre freeze) and after cryopreservation (post thaw).


RESULTS
Expression of histochemical and immunohistochemical markers as well as electron microscopy observations revealed similar adipogenic, chondrogenic and osteogenic differentiation capacity for cells isolated from pre freeze and corresponding post thaw tissue fragments of Wharton's jelly, vein or arteries of the same umbilical cord tissue, regardless growth media used for cells isolation and expansion.


CONCLUSIONS
Our efficient umbilical cord tissue cryopreservation protocol is reliable for clinical applicability of mesenchymal stem cells that could next be isolated and expanded in compliance with future accepted standards.
Regina Maria-Central Stem Cells Bank, 5B Ion Ionescu de la Brad, 13811, Bucharest, Romania; Laboratory of Histology and Developmental Biology, Faculty of Biology, University of Bucharest, Splaiul Independentei 91-95, Bucharest, R-050095, Romania.
Magda
M
Dulugiac
Department of Cellular and Molecular Biology, National Institute of Research and Development for Biological Sciences, Splaiul Independentei 296, R-060031, Bucharest, Romania.
Lucia
L
Moldovan
Laboratory of Histology and Developmental Biology, Faculty of Biology, University of Bucharest, Splaiul Independentei 91-95, Bucharest, R-050095, Romania. Electronic address: otilia.zarnescu@bio.unibuc.ro.
Otilia
O
Zarnescu
D050156:Adipogenesis; D002469:Cell Separation; D020219:Chondrogenesis; D015925:Cryopreservation; D003470:Culture Media; D005312:Fetal Blood; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000648:ultrastructure; D010012:Osteogenesis; D014470:Umbilical Cord
D003470:Culture Media
Adipogenesis; Chondrogenesis; MSCs differentiation; Osteogenesis; Umbilical cord tissue cryopreservation