Side population cells from diverse adult tissues are capable of in vitro hematopoietic differentiation

The formation of myofibroblasts in valve interstitial cell (VIC) populations contributes to fibrotic valvular disease. We examined myofibroblast differentiation in VICs from porcine aortic valves. In normal valves, cells immunostained for α-smooth muscle actin (α-SMA, a myofibroblast marker) were rare (0.69 ± 0.48%), but in sclerotic valves of animals fed an atherogenic diet, myofibroblasts were spatially clustered and abundant (31.2 ± 6.3%). In cultured VIC populations from normal valves, SMA-positive myofibroblasts were also spatially clustered, abundant (21% positive cells after 1 passage), and stained for collagen type I and vimentin but not desmin. For an analysis of stem cells, two-color flow cytometry of isolated cells stained with Hoechst 33342 demonstrated that 0.5% of VICs were side population cells; none stained for SMA. Upon culture, sorted side population cells generated ∼85% SMA-positive cells, indicating that some myofibroblasts originate from a rare population with stem cell characteristics. Plating cells on rigid collagen substrates enabled the formation of myofibroblasts after 5 days in culture, which was completely blocked by culture of cells on compliant collagen substrates. Exogenous tensile force also significantly increased SMA expression in VICs. Isotope-coded affinity tags and mass spectrometry were used to identify differentially expressed proteins in myofibroblast differentiation of VICs. Of the nine proteins that were identified, cofilin expression and phospho-cofilin were strongly increased by conditions favoring myofibroblast differentiation. Knockdown of cofilin with small-interfering RNA inhibited collagen gel contraction and reduced myofibroblast differentiation as assessed by the SMA incorporation into stress fibers. When compared with normal valves, diseased valves showed strong immunostaining for cofilin that colocalized with SMA in clustered cells. We conclude that in VICs, cofilin is a marker for myofibroblasts in vivo and in vitro that arise from a rare population of stem cells and require a rigid matrix for formation.

Download Full-text

Differentiation of CD45‑/CD31+ lung side population cells into endothelial and smooth muscle cells in vitro

International Journal of Molecular Medicine ◽

10.3892/ijmm.2019.4053 ◽

2019 ◽

Author(s):

Yang Xu ◽

Ping Sun ◽

Jian‑Yu Wang ◽

Zong‑Ze Li ◽

Rui‑Lan Gao ◽

...

Keyword(s):

Smooth Muscle ◽

Smooth Muscle Cells ◽

Side Population ◽

Muscle Cells ◽

Side Population Cells

Download Full-text

Abstract 3717: Aspirin decreases side population cells by targeting the Wnt pathway in esophageal cancer cells in vitro and enhances the combination chemotherapeutic effect of 5-FU and cisplatin in vivo.

10.1158/1538-7445.am2013-3717 ◽

2013 ◽

Author(s):

Yue Zhao ◽

Bettina Schwarz ◽

Lu Zhao ◽

Yan Wang ◽

Anneli Tischmacher ◽

...

Keyword(s):

Esophageal Cancer ◽

Cancer Cells ◽

Wnt Pathway ◽

Side Population ◽

Side Population Cells ◽

Chemotherapeutic Effect ◽

Esophageal Cancer Cells

Download Full-text

In vitro and in vivo proliferation, differentiation and migration of cardiac endothelial progenitor cells (SCA1+/CD31+ side-population cells)

Journal of Thrombosis and Haemostasis ◽

10.1111/j.1538-7836.2011.04375.x ◽

2011 ◽

Vol 9 (8) ◽

pp. 1628-1637 ◽

Cited By ~ 22

Author(s):

S. X. LIANG ◽

L. M. KHACHIGIAN ◽

Z. AHMADI ◽

M. YANG ◽

S. LIU ◽

...

Keyword(s):

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Side Population ◽

Side Population Cells ◽

Endothelial Progenitor ◽

And Migration

Download Full-text

Identification of Potential Key lncRNAs in the Context of Mouse Myeloid Differentiation by Systematic Transcriptomics Analysis

Genes ◽

10.3390/genes12050630 ◽

2021 ◽

Vol 12 (5) ◽

pp. 630

Author(s):

Yongqing Lan ◽

Meng Li ◽

Shuangli Mi

Keyword(s):

Transcription Factor ◽

Cell Model ◽

Myeloid Differentiation ◽

Rna Seq ◽

Hematopoietic Differentiation ◽

Granulocytic Differentiation ◽

Expression Of Genes ◽

Non Coding Rnas

Hematopoietic differentiation is a well-orchestrated process by many regulators such as transcription factor and long non-coding RNAs (lncRNAs). However, due to the large number of lncRNAs and the difficulty in determining their roles, the study of lncRNAs is a considerable challenge in hematopoietic differentiation. Here, through gene co-expression network analysis over RNA-seq data generated from representative types of mouse myeloid cells, we obtained a catalog of potential key lncRNAs in the context of mouse myeloid differentiation. Then, employing a widely used in vitro cell model, we screened a novel lncRNA, named Gdal1 (Granulocytic differentiation associated lncRNA 1), from this list and demonstrated that Gdal1 was required for granulocytic differentiation. Furthermore, knockdown of Cebpe, a principal transcription factor of granulocytic differentiation regulation, led to down-regulation of Gdal1, but not vice versa. In addition, expression of genes involved in myeloid differentiation and its regulation, such as Cebpa, were influenced in Gdal1 knockdown cells with differentiation blockage. We thus systematically identified myeloid differentiation associated lncRNAs and substantiated the identification by investigation of one of these lncRNAs on cellular phenotype and gene regulation levels. This study promotes our understanding of the regulation of myeloid differentiation and the characterization of roles of lncRNAs in hematopoietic system.

Download Full-text