Homo sapiens
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ABSTRACT:
PROVIDER: PRJNA170970 | ENA |
REPOSITORIES: ENA
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Project description:The fibroblast-populated 3D collagen matrix has been used to study the effect of mechanical stress on cell fate; this process is relevant to the fields of wound healing and tissue engineering. Gene array data was generated from mechanically stressed vs. stress-released matrices. The parameters of the collagen matrix model were: collagen type = bovine type I; collagen concentration = 1.5 mg/mL; initial matrix volume = 0.2 mL; initial matrix diameter = 11 mm (cultured in 24-well plates); cell type = human foreskin fibroblast, passage <10; initial matrix cell concentration = 1,000,000 cell/mL (200,000 cell/matrix); culture medium = 5% FBS in DMEM with 1 M-BM-5g/mL ascorbate. Matrices (n = 6 per experimental group) were incubated for 24 hr in the attached state; the released groups then underwent matrix detachment from the culture plate (defined as t = 0), while the attached groups were left undisturbed. RNA was isolated from attached and released matrices 6 and 24 hr after t = 0. Gene expression in the attached vs. released condition at 6 or 24 hr then was analyzed by hybridizing the anti-sense RNA derived from attached and released matrices at a given time point onto a single chip. Refer to the attached Figure 1 for the experimental design. The index experiment was defined as the comparison of gene expression in attached vs. released collagen matrices in a single strain of human foreskin fibroblasts at 6 and 24 hr after stress-release (i.e., after t = 0). Each experiment utilized two mechanical conditions (attached and released) at two time points (6 and 24 hr). So with each condition utilizing 6 matrices, each index experiment required a total of 24 matrices. In each index experiment, the chip hybridizations were: (i) 6 hr attached vs. 6 hr released, and (ii) 24 hr attached vs. 24 hr released (i.e., two gene chips per index experiment). Each hybridization was done using a 10K spotted gene chip manufactured in the UNMC Microarray Core Facility. The index experiment was performed on three fibroblast strains, meaning that expressional data was derived from three foreskin donors (nonpooled samples). Dye-swap was not performed; dye assigned to attached vs. released remained constant among all chips. Since the index experiment was performed three times, the total number of gene chips used for this entire dataset was six.
2012-07-19 | E-GEOD-39475 | biostudies-arrayexpress
Project description:The fibroblast-populated 3D collagen matrix has been used to study the effect of mechanical stress on cell fate; this process is relevant to the fields of wound healing and tissue engineering. Gene array data was generated from mechanically stressed vs. stress-released matrices. The parameters of the collagen matrix model were: collagen type = bovine type I; collagen concentration = 1.5 mg/mL; initial matrix volume = 0.2 mL; initial matrix diameter = 11 mm (cultured in 24-well plates); cell type = human foreskin fibroblast, passage <10; initial matrix cell concentration = 1,000,000 cell/mL (200,000 cell/matrix); culture medium = 5% FBS in DMEM with 1 µg/mL ascorbate. Matrices (n = 6 per experimental group) were incubated for 24 hr in the attached state; the released groups then underwent matrix detachment from the culture plate (defined as t = 0), while the attached groups were left undisturbed. RNA was isolated from attached and released matrices 6 and 24 hr after t = 0. Gene expression in the attached vs. released condition at 6 or 24 hr then was analyzed by hybridizing the anti-sense RNA derived from attached and released matrices at a given time point onto a single chip.
2012-07-20 | GSE39475 | GEO
Project description:Human Foreskin Fibroblasts-Trypanosoma cruzi infectome
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Project description:The behavior of cancer cells strongly relies on the properties of the extracellular microenvironment. The aim was to compare the whole transcriptome of three non-invasive cancer cell lines, HCT-116, A549 and T-47D, embedded in collagen I matrices in a time-dependent manner to standard two-dimensional cell culture. For this purpose, human cancer cell lines (HCT-116, A549 and T-47D) were cultured in their recommended media supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin at 37 °C, 5% CO₂ and 95% humidity. Cells were regularly tested negative for mycoplasma and authenticated by STR analysis. For 2D culture, cells were grown to 70–90% confluency and RNA was isolated. For 3D culture, single cells were embedded in 2.5 mg/mL collagen I matrices, which was polymerized at 37 °C, and cultured in medium. After 3, 5, 7, and 10 days, cells were released by collagenase IV digestion for RNA isolation.
2026-07-31 | E-MTAB-16838 | biostudies-arrayexpress
Project description:Spheroid Formation and Recovery of Human Foreskin Fibroblasts at Ambient Temperature
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