Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds
Naïve human embryonic stem cells (ESCs) are characterized by improved viability, proliferation, and differentiation capacity in comparison to traditionally derived primed human ESCs. However, currently used two-dimensional (2-D) cell culture techniques fail to mimic the three-dimensional (3-D) in vi...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-12-01
|
| Series: | Biomolecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/2218-273X/11/1/21 |
| _version_ | 1797543371996135424 |
|---|---|
| author | Christina McKee Christina Brown Shreeya Bakshi Keegan Walker Chhabi K. Govind G. Rasul Chaudhry |
| author_facet | Christina McKee Christina Brown Shreeya Bakshi Keegan Walker Chhabi K. Govind G. Rasul Chaudhry |
| author_sort | Christina McKee |
| collection | DOAJ |
| description | Naïve human embryonic stem cells (ESCs) are characterized by improved viability, proliferation, and differentiation capacity in comparison to traditionally derived primed human ESCs. However, currently used two-dimensional (2-D) cell culture techniques fail to mimic the three-dimensional (3-D) in vivo microenvironment, altering morphological and molecular characteristics of ESCs. Here, we describe the use of 3-D self-assembling scaffolds that support growth and maintenance of the naïve state characteristics of ESC line, Elf1. Scaffolds were formed via a Michael addition reaction upon the combination of two 8-arm polyethylene glycol (PEG) polymers functionalized with thiol (PEG-8-SH) and acrylate (PEG-8-Acr) end groups. 3-D scaffold environment maintained the naïve state and supported the long-term growth of ESCs. RNA-sequencing demonstrated significant changes in gene expression profiles between 2-D and 3-D grown cells. Gene ontology analysis revealed upregulation of biological processes involved in the regulation of transcription and translation, extracellular matrix organization, and chromatin remodeling in 3-D grown cells. 3-D culture conditions also induced upregulation of genes associated with Wnt and focal adhesion signaling, while p53 signaling pathway associated genes were downregulated. Our findings, for the first time, provide insight into the possible mechanisms of self-renewal of naïve ESCs stimulated by the transduction of mechanical signals from the 3-D microenvironment. |
| first_indexed | 2024-03-10T13:43:39Z |
| format | Article |
| id | doaj.art-86e394431b204664bd681ae61772339f |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-03-10T13:43:39Z |
| publishDate | 2020-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-86e394431b204664bd681ae61772339f2023-11-21T02:48:12ZengMDPI AGBiomolecules2218-273X2020-12-011112110.3390/biom11010021Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG ScaffoldsChristina McKee0Christina Brown1Shreeya Bakshi2Keegan Walker3Chhabi K. Govind4G. Rasul Chaudhry5Department of Biological Sciences, Oakland University, Rochester, MI 48309, USADepartment of Biological Sciences, Oakland University, Rochester, MI 48309, USADepartment of Biological Sciences, Oakland University, Rochester, MI 48309, USADepartment of Biological Sciences, Oakland University, Rochester, MI 48309, USADepartment of Biological Sciences, Oakland University, Rochester, MI 48309, USADepartment of Biological Sciences, Oakland University, Rochester, MI 48309, USANaïve human embryonic stem cells (ESCs) are characterized by improved viability, proliferation, and differentiation capacity in comparison to traditionally derived primed human ESCs. However, currently used two-dimensional (2-D) cell culture techniques fail to mimic the three-dimensional (3-D) in vivo microenvironment, altering morphological and molecular characteristics of ESCs. Here, we describe the use of 3-D self-assembling scaffolds that support growth and maintenance of the naïve state characteristics of ESC line, Elf1. Scaffolds were formed via a Michael addition reaction upon the combination of two 8-arm polyethylene glycol (PEG) polymers functionalized with thiol (PEG-8-SH) and acrylate (PEG-8-Acr) end groups. 3-D scaffold environment maintained the naïve state and supported the long-term growth of ESCs. RNA-sequencing demonstrated significant changes in gene expression profiles between 2-D and 3-D grown cells. Gene ontology analysis revealed upregulation of biological processes involved in the regulation of transcription and translation, extracellular matrix organization, and chromatin remodeling in 3-D grown cells. 3-D culture conditions also induced upregulation of genes associated with Wnt and focal adhesion signaling, while p53 signaling pathway associated genes were downregulated. Our findings, for the first time, provide insight into the possible mechanisms of self-renewal of naïve ESCs stimulated by the transduction of mechanical signals from the 3-D microenvironment.https://www.mdpi.com/2218-273X/11/1/21three-dimensional cultureself-assembling PEG scaffoldsnaïve embryonic stem cellssignaling pathwaysmechanotransduction |
| spellingShingle | Christina McKee Christina Brown Shreeya Bakshi Keegan Walker Chhabi K. Govind G. Rasul Chaudhry Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds Biomolecules three-dimensional culture self-assembling PEG scaffolds naïve embryonic stem cells signaling pathways mechanotransduction |
| title | Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds |
| title_full | Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds |
| title_fullStr | Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds |
| title_full_unstemmed | Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds |
| title_short | Transcriptomic Analysis of Naïve Human Embryonic Stem Cells Cultured in Three-Dimensional PEG Scaffolds |
| title_sort | transcriptomic analysis of naive human embryonic stem cells cultured in three dimensional peg scaffolds |
| topic | three-dimensional culture self-assembling PEG scaffolds naïve embryonic stem cells signaling pathways mechanotransduction |
| url | https://www.mdpi.com/2218-273X/11/1/21 |
| work_keys_str_mv | AT christinamckee transcriptomicanalysisofnaivehumanembryonicstemcellsculturedinthreedimensionalpegscaffolds AT christinabrown transcriptomicanalysisofnaivehumanembryonicstemcellsculturedinthreedimensionalpegscaffolds AT shreeyabakshi transcriptomicanalysisofnaivehumanembryonicstemcellsculturedinthreedimensionalpegscaffolds AT keeganwalker transcriptomicanalysisofnaivehumanembryonicstemcellsculturedinthreedimensionalpegscaffolds AT chhabikgovind transcriptomicanalysisofnaivehumanembryonicstemcellsculturedinthreedimensionalpegscaffolds AT grasulchaudhry transcriptomicanalysisofnaivehumanembryonicstemcellsculturedinthreedimensionalpegscaffolds |