scholarly journals Cellular Properties and Expression of Pluripotent Markers in Human Dental Pulp Stem Cells Cultured in Serum-Free Medium

2021 ◽  
Author(s):  
Chethan Kumar ◽  
Basan Gowda Sharanappa Kurkalli ◽  
Shishir Shetty ◽  
Akshay Bairapura Manjappa ◽  
Veena Shetty ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Population Doubling ◽  
Animal Origin ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Alp Activity ◽  
Human Dental Pulp

Introduction: The standard isolation and expansion of human Dental Pulp Stem Cells (DPSCs) under invitro conditions normally involve the usage of Fetal Bovine Serum (FBS). However, its animal-origin poses possible concerns for clinically relevant procedures. This critical issue compels the use of Xenogeneic-Free (XF) or human-origin alternatives to FBS for culture expansion and differentiation of DPSCs to determine the usefulness for translating into therapeutic clinical applications. Aim: To evaluate the cellular characteristics and expression of pluripotent markers in DPSCs cultured using Serum-Containing Medium (SCM-DPSCs) and Serum-Free Medium (SFM-DPSCs). Materials and Methods: This in-vitro descriptive study was conducted at NITTE (Deemed to be University), Mangaluru, Karnataka, India, from June 2019 to August 2020. DPSCs were isolated from impacted third molars. The culture expanded DPSCs in serum-containing and serum-free media were analysed on their morphology, viability, proliferation rate, Population Doubling Time (PDT), Alkaline Phosphatase (ALP) activity, cell surface markers expression, osteogenic and adipogenic potential, and the relative expression of selected pluripotent genes. Results: The primary culture of DPSCs established in SCM and SFM showed spindle shaped fibroblastic morphology with >80% viability from passage 1 (P1) to P4. A significant (p-value<0.05) difference in the proliferation rates in terms of cell numbers between SCM-DPSCs and SFM-DPSCs was observed (day 6: 3×105 vs 0.8×105; day 9: 5.8×105 vs 1.27×105; day 12: 7.8×105vs 1.56×105, respectively). The average PDT values recorded in SCM- and SFM-DPSCs were 44.33 hours and 58.41 hours, respectively. A slightly higher expression of ALP activity was observed in SCM-DPSCs than in SFM-DPSCs. Flow cytometry analysis showed that both DPSCs were positive for CD29, CD73, CD90, and negative for CD34 and CD45. The expression of OCT4 and NANOG was relatively higher in SCM-DPSCs compared to SFM-DPSCs. Further, SCM-DPSCs showed the higher levels of SOX2 and SSEA4, but did not exhibit any significant differences in their expression levels. Conclusion: The results showed that DPSCs in FBS displayed better growth kinetics and stemness markers expression along with more propensities towards lineage differentiation. SFM can be used to establish and expand DPSCs with characteristics of multipotent stem cells, but needs further research for its optimisation.

Dental pulp stem cells in serum-free medium for regenerative medicine

2019 ◽  
Vol 50 (1) ◽  
pp. 80-90 ◽  
Author(s):  
Dawn E. Coates ◽  
Mohammad Alansary ◽  
Lara Friedlander ◽  
Diogo G. Zanicotti ◽  
Warwick J. Duncan
Keyword(s):  
Stem Cells ◽  
Regenerative Medicine ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

Analysis of growth kinetics and colony-forming unit efficiency of human dental pulp stem cells cultured in different media and supplements

2021 ◽  
Vol 16 (7) ◽  
pp. 203-210
Author(s):  
Kumar Chethan ◽  
Shishir Shetty ◽  
Basan Gowda Kurkalli ◽  
Veena Shetty ◽  
Kumar Basavarajappa Mohana
Keyword(s):  
Stem Cells ◽  
Growth Kinetics ◽  
Dental Pulp ◽  
Culture Conditions ◽  
Dental Pulp Stem Cells ◽  
Population Doubling ◽  
Animal Origin ◽  
Dental Tissues ◽  
Colony Forming Unit ◽  
Human Dental Pulp

Dental tissues are considered as ideal autologous sources of multipotent stem cells. Presently, human dental pulp stem cells (DPSCs) are largely being isolated and expanded in media containing fetal bovine serum (FBS). However, the use of FBS has limitations due to its animal origin. Therefore, the present study evaluated the morphology, proliferation rate, population doubling time (PDT) and colony-forming unit fibroblast (CFU-F) efficiency of DPSCs cultured in animal serum-containing medium (SCM) and serumfree medium (SFM) in addition to serum-free culture conditions by supplementing human blood-derivatives such as platelet lysate (PL), fresh frozen plasma (FFP) and umbilical cord blood serum (UCS) at 2.5%, 5% and 7.5% concentrations. Established DPSCs had spindle-shape during primary culture but acquired characteristic fibroblast-like features when cultured in PL, FFP and UCS. DPSCs in SCM, SFM and PL had significantly (P<0.05) higher proliferative potential than those in UCS and FFP and these observations were supported by PDT values. The CFU efficiency of DPSCs was confirmed in all culture conditions with a slightly varied clonogenic potential in blood-derived components. Based on the growth kinetics and CFU ability, it is concluded that PL could be considered as a suitable alternative to FBS for the ex vivo expansion of DPSCs.

High-purity Hepatic Lineage Differentiated from Dental Pulp Stem Cells in Serum-free Medium

2012 ◽  
Vol 38 (4) ◽  
pp. 475-480 ◽  
Author(s):  
Nikolay Ishkitiev ◽  
Ken Yaegaki ◽  
Toshio Imai ◽  
Tomoko Tanaka ◽  
Taka Nakahara ◽  
...  
Keyword(s):  
Stem Cells ◽  
High Purity ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

scholarly journals Evaluation of growth, stemness, and angiogenic properties of dental pulp stem cells cultured in cGMP xeno-/serum-free medium

2019 ◽  
Vol 380 (1) ◽  
pp. 93-105 ◽  
Author(s):  
Chengjuan Qu ◽  
Maria Brohlin ◽  
Paul J Kingham ◽  
Peyman Kelk
Keyword(s):  
Stem Cells ◽  
Human Serum ◽  
Dental Pulp ◽  
Dental Pulp Stem Cells ◽  
In Vitro Assays ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cells Cultured

AbstractThis study was aimed to investigate the effects of cGMP xeno-/serum-free medium (XSF, Irvine Scientific) on the properties of human dental pulp stem cells (DPSCs). DPSCs, from passage 2, were cultured in XSF or fetal bovine serum (FBS)-supplemented medium, and sub-cultured up to passage 8. Cumulative population doublings (PDs) and the number of colony-forming-units (CFUs) were determined. qRT-PCR, ELISA, and in vitro assays were used to assess angiogenic capacity. Flow cytometry was used to measure CD73, CD90, and CD105 expression. Differentiation into osteo-, adipo-, and chondrogenic cell lineages was performed. DPSCs showed more elongated morphology, a reduced rate of proliferation at later passages, and lower CFU counts in XSF compared with FBS. Expression of angiogenic factors at the gene and protein levels varied in the two media and with passage number, but cells grown in XSF had more in vitro angiogenic activity. The majority of early and late passage DPSCs cultured in XSF expressed CD73 and CD90. In contrast, the percentage of CD105 positive DPSCs in XSF medium was significantly lower with increased passage whereas the majority of cells cultured in FBS were CD105 positive. Switching XSF-cultured DPSCs to medium supplemented with human serum restored the expression of CD105. The tri-lineage differentiation of DPSCs cultured under XSF and FBS conditions was similar. We showed that despite reduced CD105 expression levels, DPSCs expanded in XSF medium maintained a functional MSC phenotype. Furthermore, restoration of CD105 expression is likely to occur upon in vivo transplantation, when cells are exposed to human serum.

scholarly journals Characterization of human dental pulp cells grown in chemically defined serum‑free medium

2018 ◽  
Author(s):  
Sakiko Fujii ◽  
Katsumi Fujimoto ◽  
Noriko Goto ◽  
Yoshimitsu Abiko ◽  
Asayo Imaoka ◽  
...  
Keyword(s):  
Dental Pulp ◽  
Dental Pulp Cells ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Human Dental Pulp Cells ◽  
Human Dental Pulp ◽  
Pulp Cells

scholarly journals Optimizing a serum-free/xeno-free culture medium for culturing and promoting the proliferation of human dental pulp stem cells

2019 ◽  
Vol 6 ◽  
pp. 15-15 ◽  
Author(s):  
Esraa Mohamed Abdel Moniem ◽  
Mona Mahmoud EL-Batran ◽  
Ahmed Mahmoud Halawa ◽  
Dina Hazem Gomaa ◽  
Ghada Nour Eldeen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Culture Medium ◽  
Dental Pulp Stem Cells ◽  
Serum Free ◽  
Human Dental Pulp

scholarly journals In vitro induction of odontogenic activity of human dental pulp stem cells by white Portland cement enriched with zirconium oxide and zinc oxide components

2019 ◽  
Vol 13 (1) ◽  
pp. 3-10 ◽  
Author(s):  
Saeed Rahimi ◽  
Sadegh Salarinasab ◽  
Negin Ghasemi ◽  
Reza Rahbarghazi ◽  
Shahriar Shahi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Dental Pulp ◽  
Zirconium Oxide ◽  
Dental Pulp Stem Cells ◽  
Ion Release ◽  
Zno Nps ◽  
Alp Activity ◽  
Human Dental Pulp ◽  
White Portland Cement

Background. The aim of this in vitro study was to investigate the effect of zinc oxide (ZnO) and zirconium oxide (ZrO2) microparticles (MPs) and nanoparticles (NPs) in combination with white Portland cement (WPC) on odontogenic capacity of human dental pulp stem cells over a period of 21 days. Methods. Synthesized ZnO and ZrO2 particles were characterized using scanning electron microscopy and transmission electron microscopy. The viability of human dental pulp stem cells was measured by a 3-(4,5-dimethylthiazolyl-2-yl)-2,5- diphenyltetrazolium bromide assay at 7-, 14- and 21-day intervals after seeding on WPC disks enriched with ZnO and ZrO2 MPs and NPs. Odontogenic potential of ZnO and ZrO2 particles in combination with WPC was investigated by alkaline phosphatase (ALP) activity and ionized calcium level of supernatant culture media at different time intervals. Data were analyzed using one-way ANOVA and post hoc Tukey tests. Results. All the materials exhibited cell viability over a 21-day period, except for WPC with ZnO NPs on day 7, although it was not statistically significant (P>0.05). The ALP activity and ionized calcium level increased in all the groups compared to the control group (P<0.05). ZnO NPs had superior effect on odontogenic activity and calcium ion release compared to ZnO MPs (P=0.046). There was no significant difference between ZrO2 MPs and NPs in odontogenic activity (P>0.05). Conclusion. WPC enriched with ZnO and ZrO2 increased ALP activity and calcium ion release of human dental pulp stem cells over a period of 21 days in vitro.

Sign in / Sign up

Export Citation Format

Share Document