Fourier transform infrared microspectroscopy reveals that tissue culture conditions affect the macromolecular phenotype of human embryonic stem cells

The Analyst ◽  
2013 ◽  
Vol 138 (14) ◽  
pp. 4147 ◽  
Author(s):  
Julie Cao ◽  
Elizabeth S. Ng ◽  
Don McNaughton ◽  
Edouard G. Stanley ◽  
Andrew G. Elefanty ◽  
...  
Keyword(s):  
Stem Cells ◽  
Tissue Culture ◽  
Fourier Transform ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Fourier Transform Infrared ◽  
Culture Conditions ◽  
Infrared Microspectroscopy ◽  
Fourier Transform Infrared Microspectroscopy

scholarly journals Morphogen And Community Effects Determine Cell Fates In Response To BMP4 Signaling In Human Embryonic Stem Cells

2017 ◽  
Author(s):  
Anastasiia Nemashkalo ◽  
Albert Ruzo ◽  
Idse Heemskerk ◽  
Aryeh Warmflash
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Cell Fate ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Culture Conditions ◽  
Community Effects ◽  
Cell Fates ◽  
Standard Culture

AbstractParacrine signals maintain developmental states and create cell-fate patterns in vivo, and influence differentiation outcomes in human embryonic stem cells (hESCs) in vitro. Systematic investigation of morphogen signaling is hampered by the difficulty of disentangling endogenous signaling from experimentally applied ligands. Here, we grow hESCs in micropatterned colonies of 1-8 cells (“μColonies”) to quantitatively investigate paracrine signaling and the response to external stimuli. We examine BMP4-mediated differentiation in μColonies and standard culture conditions and find that in μColonies, above a threshold concentration, BMP4 gives rise to only a single cell fate, contrary to its role as a morphogen in other developmental systems. Under standard culture conditions, BMP4 acts as morphogen, but this effect requires secondary signals and particular cell densities. We further find that a “community effect” enforces a common fate within μColonies both in the state of pluripotency and when cells are differentiated, and that this effect allows more precise response to external signals. Using live cell imaging to correlate signaling histories with cell fates, we demonstrate that interactions between neighbors result in sustained, homogenous signaling necessary for differentiation.Summary StatementWe quantitatively examined signaling and differentiation in hESC colonies of varying size treated with BMP4. We show that secondary signals result in morphogen and community effects that determine cell fates.

scholarly journals 176 IMPROVED GROWTH OF HUMAN EMBRYONIC STEM CELLS IN A REDUCED OXYGEN ATMOSPHERE

2005 ◽  
Vol 17 (2) ◽  
pp. 238 ◽  
Author(s):  
T. Peura ◽  
A. Bosman ◽  
T. Stojanov
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Culture Conditions ◽  
Oxygen Atmosphere ◽  
Low Oxygen ◽  
Treatment Groups ◽  
Absolute Growth ◽  
Differentiation Status

Traditional cell culture conditions entail the use of gas atmosphere consisting of 5–6% CO2 in air. These same conditions have also been used universally for the culture of human embryonic stem cells (hES), despite the natural milieu of the embryos, from which these cells are derived, being slightly hypoxic. The aim of this work was to examine if human embryonic stem cells would benefit from the reduced oxygen culture environment, as used for human pre-implantation embryos. In Experiment 1, the relative growth in high and low oxygen atmosphere was compared by cutting undifferentiared hES-colonies into equal-size pieces and transferring them to two dishes, one into the conventional incubator in 6% CO2 in air (HIGH), the other into K-MINC-1000 (Eight Mile Plains, Queensland) mini-incubator in 5% O2, 6% CO2 and 89% N2 (LOW). After 8 days the colony sizes and differentiation status were measured. In Experiment 2, the absolute growth in high and low oxygen atmosphere was compared by cutting exactly same size fragments from undifferentiated hES-colonies and distributing them to two treatments as described. All colonies were measured and passaged at Day 7 and final measurements taken at Day 14. In Experiment 1, improved growth was observed in reduced oxygen, mean percentage of undifferentiated growth from original colony (assigned as 100%) being 217% for HIGH and 482% for LOW over three replicates. In Experiment 2, the total of 16.9 mm2 of undifferentiated colonies in both treatment groups (24 colonies per group over 3 replicates) had by Day 7 grown to total of 51.1 and 79.5 mm2 (P < 0.001), and by Day 14 to 216.8 and 373.3 mm2 (P < 0.0001) in HIGH and LOW, respectively (see Table 1). In neither experiment were there differences in the differentiation status of the colonies between the treatments (mean 6% and 5% at Day 7, and 9% and 9% at Day 14 in HIGH and LOW, respectively). We conclude that culture in reduced oxygen improves growth of human embryonic stem cells. Table 1. Growth of hES colonies on high and low oxygen atmosphere (data from 3 replicates) We like to acknowledge Bresagen Ltd. for the supply of hesBGN-01 cell line.

Adaptation of human embryonic stem cells to feeder-free and matrix-free culture conditions directly on plastic surfaces

2008 ◽  
Vol 133 (1) ◽  
pp. 146-153 ◽  
Author(s):  
Narmin Bigdeli ◽  
Maria Andersson ◽  
Raimund Strehl ◽  
Katarina Emanuelsson ◽  
Eva Kilmare ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Culture Conditions ◽  
Plastic Surfaces ◽  
Matrix Free
Sign in / Sign up

Export Citation Format

Share Document