Culture and molecular markers characterization of Stem Cells from Human Deciduous (SHED) and Permanent (DPSC) teeth pulp

Shahrul Hisham Zainal Ariffin, Thanaletchumi Manogaran, Intan Zarina Zainol Abidin, Zaidah Zainal Ariffin, Pusat Pengajian Biologi, Rohaya Megat Abdul Wahab

Research output: Contribution to journalArticle

Abstract

Human dental pulp stem cells are adult multipotent stem cells isolated from dental pulp tissue. Our objective was to determine in vitro culture technique for stem cells from deciduous tooth (SHED) and permanent tooth (DPSC) through cell passage identification, the effect of trypsin-EDTA and proliferation potential, and to characterize both cells using molecular markers profile. Enzyme digestion method was used on dental pulp tissue from deciduous and permanent tooth for SHED and DPSC isolation, respectively. Both cells were cultured at passage 1 until 5 and trypan blue assay was used to obtain growth curve in determining cell population doubling time (PDT) for each passage. Effect of trypsin-EDTA on both cells were studied using Alamar blue assay to determine optimum incubation time for subculturing process. Cell proliferation potential for both cells within 21 days was determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cell characterization through molecular biology markers was performed using RT-PCR. Both cells at all passages appeared fibroblast-like. SHED and DPSC at passage 3 exhibited the lowest PDT with 43 ± 2.3 and 63 ± 3.1 h, respectively. SHED exposed to trypsin-EDTA showed decrease in cell viability percentage compared to DPSC. Cell growth for SHED was ∼2.3-fold higher than DPSC. Both cells expressed mesenchymal stem cell markers and not hematopoietic stem cell markers. In conclusion, homogenous morphology and lowest PDT value indicated that cells at passage 3 are the best to determine proliferation potential and molecular markers expression. SHED proliferated better than DPSC. However, DPSC was more resistant to trypsin-EDTA than SHED. Based on molecular marker profile, both cells are mesenchymal stem cells.

Original languageEnglish
Pages (from-to)1855-1865
Number of pages11
JournalSains Malaysiana
Volume48
Issue number9
DOIs
Publication statusPublished - 1 Jan 2019

Fingerprint

Tooth
Stem Cells
Edetic Acid
Dental Pulp
Trypsin
Deciduous Tooth
Mesenchymal Stromal Cells
Multipotent Stem Cells
Population
Culture Techniques
Adult Stem Cells
Trypan Blue
Growth
Hematopoietic Stem Cells
Molecular Biology
Digestion
Cultured Cells
Cell Survival
Fibroblasts
Cell Proliferation

Keywords

  • Cell doubling time
  • Human dental pulp stem cells
  • Molecular biology markers profile
  • Proliferation potential
  • Trypsin-EDTA effect

ASJC Scopus subject areas

  • General

Cite this

Culture and molecular markers characterization of Stem Cells from Human Deciduous (SHED) and Permanent (DPSC) teeth pulp. / Ariffin, Shahrul Hisham Zainal; Manogaran, Thanaletchumi; Abidin, Intan Zarina Zainol; Ariffin, Zaidah Zainal; Pengajian Biologi, Pusat; Wahab, Rohaya Megat Abdul.

In: Sains Malaysiana, Vol. 48, No. 9, 01.01.2019, p. 1855-1865.

Research output: Contribution to journalArticle

Ariffin, Shahrul Hisham Zainal ; Manogaran, Thanaletchumi ; Abidin, Intan Zarina Zainol ; Ariffin, Zaidah Zainal ; Pengajian Biologi, Pusat ; Wahab, Rohaya Megat Abdul. / Culture and molecular markers characterization of Stem Cells from Human Deciduous (SHED) and Permanent (DPSC) teeth pulp. In: Sains Malaysiana. 2019 ; Vol. 48, No. 9. pp. 1855-1865.
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