Proliferation and osteoblast differentiation mice dental pulp stem cells between enzyme digestion and outgrowth method

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Abstract

The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential of mice DPSCs into bone cells. Dental pulp was extracted from mice's incisors and subjected to isolation either by collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblastliked morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation. Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.

Original languageEnglish
Pages (from-to)691-698
Number of pages8
JournalSains Malaysiana
Volume47
Issue number4
DOIs
Publication statusPublished - 1 Apr 2018

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Dental Pulp
Osteoblasts
Digestion
Stem Cells
Enzymes
Eagles
Cell Separation
Collagenases
Incisor
Alkaline Phosphatase
Bone and Bones

Keywords

  • Adherent cells
  • Alkaline phosphatase
  • In vitro
  • Mesenchymal stem cells

ASJC Scopus subject areas

  • General

Cite this

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abstract = "The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential of mice DPSCs into bone cells. Dental pulp was extracted from mice's incisors and subjected to isolation either by collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblastliked morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation. Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.",
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author = "Farinawati Yazid and Luchman, {Nur Atmaliya} and {Megat Abdul Wahab}, Rohaya and {Zainal Ariffin}, {Shahrul Hisham} and Sahidan Senafi",
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AU - Yazid, Farinawati

AU - Luchman, Nur Atmaliya

AU - Megat Abdul Wahab, Rohaya

AU - Zainal Ariffin, Shahrul Hisham

AU - Senafi, Sahidan

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N2 - The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential of mice DPSCs into bone cells. Dental pulp was extracted from mice's incisors and subjected to isolation either by collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblastliked morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation. Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.

AB - The isolation method for dental pulp stem cells (DPSCs) is still unclear to obtain a conducive environment for DPSCs to proliferate. Enzymatic digestion and outgrowth method are two commonly used methods for DPSCs isolation but are not well characterized in mice DPSCs. This study aimed to compare these isolation methods and differentiation potential of mice DPSCs into bone cells. Dental pulp was extracted from mice's incisors and subjected to isolation either by collagenase 1A or culture of pulp tissue in complete alpha-Modified Eagle Medium (αMEM). Both cells isolated were cultured until passage 4 and subjected to in vitro proliferation and differentiation analysis. Both cells exhibited fibroblastliked morphology, but cells isolated by enzyme digestion proliferate faster compare to outgrowth method. After 21 days of osteoblast differentiation, DPSCs isolated from enzyme digestion method showed alkaline phosphatase (ALP) activity slightly different as compared to outgrowth method. In conclusion, there is a significant difference between the cells isolated from enzyme digestion compare to outgrowth method with regard to proliferation and osteoblast differentiation. Thus, it is preferable to isolate by enzyme digestion as it is faster and consistent compared to outgrowth method.

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