Chareerut PhruksaniyomThanasup GonmaneeKutkao VongsavanTaweepong ArayapisitPetcharat KraivaphanHathaitip SritanaudomchaiMahidol University2018-12-212019-03-142018-12-212019-03-142017-01-01Journal of Stem Cells. Vol.12, No.4 (2017), 161-173155685392-s2.0-85045880345https://repository.li.mahidol.ac.th/handle/20.500.14594/41994© Nova Science Publishers, Inc. Stem cells from human exfoliated deciduous teeth (SHED) are originally referred to as fibroblastoid-colony-forming-cells because one of their characteristics is related to proliferation and differentiation potential. In this study, we developed an efficient protocol for colony-forming-unit (CFU) assay for SHED. The culture conditions including culturing time, seeding cell density and modifications of media supplements were optimized. The CFUs were determined on the proportion of colony types, sizes and the numbers of colony. Isolated SHED expressed CD44, CD73, CD105 and showed multipotent ability. Higher yield was obtained when SHED was cultured at the density of 500 cells per well of a 6-well plate for 14 days in Dulbecco’s modified Eagle medium (DMEM) with 10% fetal bovine serum (FBS). The number and size of CFU was increased with the increase of FBS concentration in which the transit-amplifying cell (TAC) colonies were the most abundant, followed by the stem cell (SC) and differentiated cell (DC) colonies, respectively. In DMEM with 20% FBS and 100 µM L-ascorbic acid-2-phosphate, the CFU rate was highest with increasing the proportion of SC colonies. The improvement of the culture conditions described herein could be adopted to easily acquire more functional SHED for applying in cell therapy.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS