Three-dimensional culture of dental pulp pluripotent-like stem cells (DPPSCs) enhances Nanog expression and provides a serum-free condition for exosome isolation

FASEB BioAdvances
Farid N FaruquKhuloud T Al-Jamal

Abstract

Stem cell-derived exosomes have been identified as novel cell-free therapeutics for regenerative medicine. Three-dimensional (3D) culture of stem cells were reported to improve their "stemness" and therapeutic efficacy. This work focused on establishing serum-free 3D culture of dental pulp pluripotent-like stem cells (DPPSCs)-a newly characterized pluripotent-like stem cell for exosome production. DPPSCs were expanded in regular 2D culture in human serum-supplemented (HS)-medium and transferred to a micropatterned culture plate for 3D culture in HS-medium (default) and medium supplemented with KnockOut™ serum replacement (KO-medium). Bright-field microscopy observation throughout the culture period (24 days) revealed that DPPSCs in KO-medium formed spheroids of similar morphology and size to that in HS-medium. qRT-PCR analysis showed similar Oct4A gene expression in DPPSC spheroids in both HS-medium and KO-medium, but Nanog expression significantly increased in the latter. Vesicles isolated from DPPSC spheroids in KO-medium in the first 12 days of culture showed sizes that fall within the exosomal size range by nanoparticle tracking analysis (NTA) and express the canonical exosomal markers. It is concluded that 3D culture of DP...Continue Reading

References

Nov 14, 1997·The American Journal of Physiology·W Mueller-Klieser
Sep 2, 1999·The New England Journal of Medicine·A J Singer, R A Clark
Sep 6, 2001·Proceedings of the National Academy of Sciences of the United States of America·D S KaufmanJ A Thomson
May 4, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Jeffrey L SpeesDarwin J Prockop
Apr 15, 2005·Genesis : the Journal of Genetics and Development·Rie ShimizukawaKen-ichi Yagami
May 24, 2005·International Immunology·Marie-Pierre CabyChristian Bonnerot
Sep 27, 2005·Alternatives to Laboratory Animals : ATLA·Sandra CoeckeUNKNOWN Second ECVAM Task Force on Good Cell Culture Practice
Apr 19, 2006·Journal of Cellular Biochemistry·Arnold I Caplan, James E Dennis
Jun 8, 2007·Nature·Keisuke OkitaShinya Yamanaka
Jul 5, 2007·Journal of Bioscience and Bioengineering·Hiroshi Kurosawa
Nov 9, 2007·Tissue Engineering·Wesley L RandleAthanasios Mantalaris
Nov 22, 2007·Science·Junying YuJames A Thomson
Nov 27, 2007·Journal of Tissue Engineering and Regenerative Medicine·Abdallah S Daar, Heather L Greenwood
May 16, 2008·Nature·Geoffrey C GurtnerMichael T Longaker
May 22, 2008·Journal of Tissue Engineering and Regenerative Medicine·Stefan BajadaNureddin Ashammakhi
Oct 14, 2008·Nature Biotechnology·Danwei HuangfuDouglas A Melton
Dec 5, 2008·Journal of Tissue Engineering and Regenerative Medicine·Juan ZhangAthanasios Mantalaris
Feb 7, 2009·Biotechnology Progress·Andrés M Bratt-LealTodd C McDevitt
Feb 21, 2009·Journal of Tissue Engineering and Regenerative Medicine·Rakhi PalSatish Totey
Apr 25, 2009·Journal of the American Society of Nephrology : JASN·Stefania BrunoGiovanni Camussi
May 14, 2009·Transplantation·Georg SiegelFrancesco Dazzi
Aug 26, 2009·Cell·Jose SilvaAustin Smith
Feb 9, 2010·Stem Cell Research·Ruenn Chai LaiSai Kiang Lim
Apr 7, 2010·Toxicology in Vitro : an International Journal Published in Association with BIBRA·J van der ValkG Gstraunthaler
Jul 6, 2010·Journal of Proteomics·Suresh MathivananRichard J Simpson
Jul 21, 2010·Proceedings of the National Academy of Sciences of the United States of America·Thomas J BartoshDarwin J Prockop
Jan 1, 1990·Toxicology in Vitro : an International Journal Published in Association with BIBRA·M Taub
Feb 23, 2011·The Journal of Pathology·Shidou ZhaoAijun Hao

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Methods Mentioned

BETA
PCR
dot blot

Software Mentioned

Primer
Image [UNK]
ImageJ
7500
BLAST
OligoAnalyzer
NanoSight NTA

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