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Stem Cell Research and Therapy

, 8:139

First Online: 08 June 2017Received: 21 November 2016Revised: 24 April 2017Accepted: 22 May 2017DOI: 10.1186-s13287-017-0594-2

Cite this article as: Kim, Y.M., Kang, Y.G., Park, S.H. et al. Stem Cell Res Ther 2017 8: 139. doi:10.1186-s13287-017-0594-2

Abstract

BackgroundMechanical stimuli play important roles in the proliferation and differentiation of adult stem cells. However, few studies on their effects on induced pluripotent stem cells iPSCs have been published.

MethodsHuman dermal fibroblasts were seeded onto flexible membrane-bottom plates, and infected with retrovirus expressing the four reprogramming factors OCT4, SOX2, KLF, and c-MYC OSKM. The cells were subjected to equiaxial stretching 3% or 8% for 2, 4, or 7 days and seeded on feeder cells STO. The reprogramming into iPSCs was evaluated by the expression of pluripotent markers, in vitro differentiation into three germ layers, and teratoma formation.

ResultsEquiaxial stretching enhanced reprogramming efficiency without affecting the viral transduction rate. iPSCs induced by transduction of four reprogramming factors and application of equiaxial stretching had characteristics typical of iPSCs in terms of pluripotency and differentiation potentials.

ConclusionsThis is the first study to show that mechanical stimuli can increase reprogramming efficiency. However, it did not enhance the infection rate, indicating that mechanical stimuli, defined as stretching in this study, have positive effects on reprogramming rather than on infection. Additional studies should evaluate the mechanism underlying the modulation of reprogramming of somatic cells into iPSCs.

KeywordsHuman-induced pluripotent stem cells Cell reprogramming Reprogramming factors Human dermal fibroblasts Mechanical stimulation Equiaxial stretching AbbreviationsALPAlkaline phosphatase

ANOVAAnalysis of variance

bFGFBasic fibroblast growth factor

BSABovine serum albumin

CPCommercial plates

DDays

DMEM-F12Dulbecco’s modified Eagle’s medium-Ham’s F-12

DMEMDulbecco’s modified Eagle’s medium

EBEmbryoid body

EDTAEthylenediaminetetraacetic acid

ESEmbryonic stem

EtBrEthidium bromide

FACSFluorescence-activated cell sorting

FBSFetal bovine serum

FGM-2Fibroblast growth medium-2

FMFlexible membranes

GFPGreen fluorescent protein

HandEHematoxylin and eosin

iPSCInduced pluripotent stem cell

KLF4Kruppel-like factor 4

NEAANon-essential amino acids

OCT4Octamer-binding transcription factor 4

ONSLOCT4, NANOG, SOX2, and Lin28

OSKMOCT4, SOX2, KLF4, and c-MYC

PBSPhosphate-buffered saline

PVDFPolyvinylidene fluoride

Retro-OSKMpMXs-OCT4, pMXs-SOX2, pMXs-KLF, pMXs-c-MYC

RT-PCRReverse-transcription polymerase chain reaction

SOX2Sex determining region Y-box 2

SRKnockout serum replacement

VPAValproic acid





Autor: Young Mi Kim - Yun Gyeong Kang - So Hee Park - Myung-Kwan Han - Jae Ho Kim - Ji Won Shin - Jung-Woog Shin

Fuente: https://link.springer.com/







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