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Pooled Human Serum Increases Regenerative Potential of In Vitro Expanded Stem Cells from Human Extracted Deciduous Teeth

  • Nazmul Haque
  • Noor Hayaty Abu Kasim
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1083)

Abstract

In regenerative therapy, in vitro expansion of stem cells is critical to obtain a significantly higher number of cells for successful engraftment after transplantation. However, stem cells lose its regenerative potential and enter senescence during in vitro expansion. In this study, the influence of foetal bovine serum (FBS) and pooled human serum (pHS) on the proliferation, morphology and migration of stem cells from human extracted deciduous teeth (SHED) was compared. SHED (n = 3) was expanded in KnockOut DMEM supplemented with either pHS (pHS-SM) or FBS (FBS-SM). pHS was prepared using peripheral blood serum of six healthy male adults, aged between 21 and 35 years old. The number of live SHED was significantly higher, from passage 5 to 7, when cultured in pHS-SM compared to those cultured in FBS-SM (p < 0.05). Number of cells having flattened morphology, characteristics of partially differentiated and senescent cells, was significantly lower (p < 0.05) in pHS-SM (3%) compared to those in FBS-SM (7%). Furthermore, migration of SHED in pHS-SM was found to be more directional. The presence of selected ten paracrine factors known for their proliferation and migration potential was detected in all six individual human sera, used to produce pHS, none of which were detected in FBS. Ingenuity Pathway Analysis showed the possible involvement of the ‘ephrin receptor signalling pathway’ to regulate the proliferation and migration of SHED in pHS-SM. In conclusion, pHS-SM showed significantly higher proliferation rate and could maintain significantly lower number of senescent cells and support directional migration of cells.

Keywords

Engraftment Foetal bovine serum Morphology Paracrine factors Regenerative medicine 

Abbreviations

BM

bone marrow

EGF

epidermal growth factor

FBS

foetal bovine serum

FC

flattened

FGF-2

fibroblast growth factor-2

FSS

flat spindle-shaped

G-CSF

granulocyte colony-stimulating factor

GM-CSF

granulocyte macrophage colony-stimulating factor

HGF

hepatocyte growth factor

LIF

leukaemia inhibitory factor

MSCs

mesenchymal stem cells

PDGF-BB

platelet-derived growth factor BB

pHS

pooled human serum

RS

rapidly self-renewing

SCF

stem cell factor

SDF-1α

stromal cell-derived factor-1α

SHED

stem cells from human extracted deciduous teeth

SS

spindle-shaped

VEGF

vascular endothelial growth factor

Notes

Acknowledgement

This work was supported by High Impact Research MOHE Grant UM.C/625/1/HIR/MOHE/DENT/01 from the Ministry of Higher Education Malaysia. The authors thank the staff nurses from Oral and Maxillofacial Sciences Department, Faculty of Dentistry, University of Malaya, for their support in the collection of blood from donors.

Conflicts of Interest

The authors deny any conflicts of interest related to this study.

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© Springer International Publishing AG  2017

Authors and Affiliations

  1. 1.Department of Restorative Dentistry, Faculty of DentistryUniversity of MalayaKuala LumpurMalaysia
  2. 2.Regenerative Dentistry Research Group, Faculty of DentistryUniversity of MalayaKuala LumpurMalaysia

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