Stem Cells from Human Exfoliated Deciduous Teeth – Isolation, Long Term Cultivation and Phenotypical Analysis

Suchánek, Jakub; Víšek, Benjamín; Soukup, Tomáš; El-Din Mohamed, Sally Kamal; Ivančaková, Romana; Mokrý, Jaroslav; Aboul-Ezz, Eman H. A.; Omran, A.

doi:10.14712/18059694.2016.66

Acta Medica > Archive > 2010, Vol. 53 > Issue 2 > Stem Cells from Human Exfoliated…

Acta Med. 2010, 53: 93-99

https://doi.org/10.14712/18059694.2016.66

Stem Cells from Human Exfoliated Deciduous Teeth – Isolation, Long Term Cultivation and Phenotypical Analysis

Jakub Suchánek^a, Benjamín Víšek^b, Tomáš Soukup^b, Sally Kamal El-Din Mohamed^c, Romana Ivančaková^a, Jaroslav Mokrý^b, Eman H. A. Aboul-Ezz^c, A. Omran^d

^aCharles University in Prague, Faculty of Medicine and University Hospital Hradec Králové, Department of Dentistry, Hradec Králové, Czech Republic
^bCharles University in Prague, Faculty of Medicine and University Hospital Hradec Králové, Department of Histology and Embryology, Hradec Králové, Czech Republic
^cOro-Dental Genetic Department, National Research Center, Cairo, Egypt
^dFaculty of Dentistry, Ein-Shams University, Cairo, Egypt

Received February 8, 2010
Accepted May 6, 2010

Aims: Our aims were to isolate stem cells from human exfoliated deciduous teeth (SHED), to cultivate them in vitro and to investigate their basic biological properties, phenotype and to compare our findings with dental pulp stem cells (DPSC) isolated from permanent teeth. Methods: Dental pulp was gently evacuated from exfoliated teeth. After enzymatic dissociation of dental pulp, SHED were cultivated in modified cultivation media for mesenchymal adult progenitor cells containing 2 % FCS and supplemented with growth factors and insulin, transferrin, sodium (ITS) supplement. Cell viability and other biological properties were examined using a Vi-Cell analyzer and a Z2-Counter. DNA analyses and phenotyping were performed with flow cytometry. Results: We were able to cultivate SHED over 45 population doublings. Our results showed that SHED cultivated under same conditions as DPSC had longer average population doubling time (41.3 hrs for SHED vs. 24.5 hrs for DPSC). Phenotypic comparison of cultivated SHED to that of cultivated DPSC showed differential expression CD29, CD44, CD71, CD117, CD166. During long-term cultivation, SHED did not showed any signs of degeneration or spontaneous differentiation. Conclusions: We isolated stem cells from exfoliated teeth. In comparison to DPSC, SHED proliferation rate was about 50% slower, and SHED showed slightly different phenotype. These cells may be extremely useful for stem cell tissue banking, further stem cell research and future therapeutic applications.