Culture and Molecular Markers Characterization of Stem Cells from Human Deciduous (SHED) and Permanent (DPSC) Teeth Pulp

被引:3
作者
Ariffin, Shahrul Hisham Zainal [1 ]
Manogaran, Thanaletchumi [1 ]
Abidin, Intan Zarina Zaino [2 ]
Ariffin, Zaidah Zainal [3 ]
Wahab, Rohaya Megat Abdul [4 ]
机构
[1] Univ Kebangsaan Malaysia, Fak Sains & Teknol, Pusat Bioteknol & Makanan Berfungsi, Bangi 43600, Selangor Darul, Malaysia
[2] Cyberjaya Univ, Ctr Res & Postgrad Studies, Coll Med Sci, Cyberjaya 63000, Selangor Darul, Malaysia
[3] Univ Teknol MARA, Fak Sains Gunaan, Pusat Pengajian Biol, Uitm Shah Alam 40450, Selangor Darul, Malaysia
[4] Univ Kebangsaan Malaysia, Fak Pergigian, Pusat Kesihatan Pergigian Keluarga, Unit Ortodont, Kuala Lumpur 50300, Wilayah Perseku, Malaysia
来源
SAINS MALAYSIANA | 2019年 / 48卷 / 09期
关键词
Cell doubling time; human dental pulp stem cells; molecular biology markers profile; proliferation potential; trypsin-EDTA effect; DENTAL-PULP; DERMAL FIBROBLASTS; CONDITIONED MEDIUM; IN-VITRO; EXPRESSION; KERATINOCYTES; EXPANSION; CAPACITY; CD105; CD73;
D O I
10.17576/jsm-2019-4809-06
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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)-25-diphenyltetrazolium bromide (AITT) 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 nypsin-EDTA showed decrease in cell viability percentage compared to DPSC. Cell growth for SHED was similar to 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.
引用
收藏
页码:1855 / 1865
页数:11
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