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Employment of gene expression profiling to identify transcriptional regulators of hepatic stellate cells.

Shimada H, Rajagopalan LE - Fibrogenesis Tissue Repair (2012)

Bottom Line: The molecular mechanisms underlying this process are not fully understood.Examination of microarray data following Matrigel-induced deactivation of HSC revealed a significant down-regulation of myocardin, an important transcriptional regulator in smooth and cardiac muscle development.Thus, gene expression profiling as well as functional assays of activated HSC have provided the first evidence of the involvement of myocardin in HSC activation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Inflammation Research Unit, Pfizer Global Research and Development, Pfizer Inc, 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA.

ABSTRACT
Activated hepatic stellate cells (HSC) play a central role in scar formation that leads to liver fibrosis. The molecular mechanisms underlying this process are not fully understood. Microarray and bioinformatics analyses have proven to be useful in identifying transcription factors that regulate cellular processes such as cell differentiation. Using oligonucleotide microarrays, we performed transcriptional analyses of activated human HSC cultured on Matrigel-coated tissue culture dishes. Examination of microarray data following Matrigel-induced deactivation of HSC revealed a significant down-regulation of myocardin, an important transcriptional regulator in smooth and cardiac muscle development. Thus, gene expression profiling as well as functional assays of activated HSC have provided the first evidence of the involvement of myocardin in HSC activation.

No MeSH data available.


Related in: MedlinePlus

Effect of Matrigel-induced deactivation on the morphology of activated rat activated HSCs. Primary rat HSCs were plated either on plastic (left) or Matrigel-coated (right) tissue culture dishes and cultured for 3 days. Confocal microscopic images of HSC plated on plastic or Matrigel-coated dishes following staining with α-SMA antibody (green) and TOPRO-3 nuclear stain (blue). HSCs cultured on plastic dishes exhibit myofibroblast-like features, immunostaining for α-SMA, while cells plated on Matrigel-coated dishes stained weakly for α-SMA.
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Figure 1: Effect of Matrigel-induced deactivation on the morphology of activated rat activated HSCs. Primary rat HSCs were plated either on plastic (left) or Matrigel-coated (right) tissue culture dishes and cultured for 3 days. Confocal microscopic images of HSC plated on plastic or Matrigel-coated dishes following staining with α-SMA antibody (green) and TOPRO-3 nuclear stain (blue). HSCs cultured on plastic dishes exhibit myofibroblast-like features, immunostaining for α-SMA, while cells plated on Matrigel-coated dishes stained weakly for α-SMA.

Mentions: The ECM components regulate cellular process such as cell shape, motility, growth, differentiation and gene expression. BD Matrigel, a basement membrane matrix marketed by BD Biosciences is a gelatinous protein mixture secreted by Engelbroth-Hom-Swarm (EHS) mouse sarcoma cells and resembles the complex extracellular matrix environment. When cultured on plastic dishes, isolated quiescent HSC spontaneously transform to the activated state. However, culturing on Matrigel-coated dishes maintains the cells in a quiescent state [16]. Additionally, activated HSC can be deactivated on Matrigel, resulting in decreased α-SMA and collagen gene expression [17,18]. Figure 1. shows a comparison of rat primary HSC cultured either on plastic or Matrigel-coated dishes for 3 days, with Matrigel cultured stellate cells exhibiting a significantly reduced expression of α-SMA. Similarly, human HSC cell lines, LI90 and LX-2 cultured on Matrigel-coated dishes were deactivated, taking on around, compact appearance. Even though it is still unclear whether Matrigel can induce HSC reversion to a quiescent state, the cells cultured on Matrigel-coated dish were apparently deactivated.


Employment of gene expression profiling to identify transcriptional regulators of hepatic stellate cells.

Shimada H, Rajagopalan LE - Fibrogenesis Tissue Repair (2012)

Effect of Matrigel-induced deactivation on the morphology of activated rat activated HSCs. Primary rat HSCs were plated either on plastic (left) or Matrigel-coated (right) tissue culture dishes and cultured for 3 days. Confocal microscopic images of HSC plated on plastic or Matrigel-coated dishes following staining with α-SMA antibody (green) and TOPRO-3 nuclear stain (blue). HSCs cultured on plastic dishes exhibit myofibroblast-like features, immunostaining for α-SMA, while cells plated on Matrigel-coated dishes stained weakly for α-SMA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3368757&req=5

Figure 1: Effect of Matrigel-induced deactivation on the morphology of activated rat activated HSCs. Primary rat HSCs were plated either on plastic (left) or Matrigel-coated (right) tissue culture dishes and cultured for 3 days. Confocal microscopic images of HSC plated on plastic or Matrigel-coated dishes following staining with α-SMA antibody (green) and TOPRO-3 nuclear stain (blue). HSCs cultured on plastic dishes exhibit myofibroblast-like features, immunostaining for α-SMA, while cells plated on Matrigel-coated dishes stained weakly for α-SMA.
Mentions: The ECM components regulate cellular process such as cell shape, motility, growth, differentiation and gene expression. BD Matrigel, a basement membrane matrix marketed by BD Biosciences is a gelatinous protein mixture secreted by Engelbroth-Hom-Swarm (EHS) mouse sarcoma cells and resembles the complex extracellular matrix environment. When cultured on plastic dishes, isolated quiescent HSC spontaneously transform to the activated state. However, culturing on Matrigel-coated dishes maintains the cells in a quiescent state [16]. Additionally, activated HSC can be deactivated on Matrigel, resulting in decreased α-SMA and collagen gene expression [17,18]. Figure 1. shows a comparison of rat primary HSC cultured either on plastic or Matrigel-coated dishes for 3 days, with Matrigel cultured stellate cells exhibiting a significantly reduced expression of α-SMA. Similarly, human HSC cell lines, LI90 and LX-2 cultured on Matrigel-coated dishes were deactivated, taking on around, compact appearance. Even though it is still unclear whether Matrigel can induce HSC reversion to a quiescent state, the cells cultured on Matrigel-coated dish were apparently deactivated.

Bottom Line: The molecular mechanisms underlying this process are not fully understood.Examination of microarray data following Matrigel-induced deactivation of HSC revealed a significant down-regulation of myocardin, an important transcriptional regulator in smooth and cardiac muscle development.Thus, gene expression profiling as well as functional assays of activated HSC have provided the first evidence of the involvement of myocardin in HSC activation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Inflammation Research Unit, Pfizer Global Research and Development, Pfizer Inc, 700 Chesterfield Parkway West, Chesterfield, MO 63017, USA.

ABSTRACT
Activated hepatic stellate cells (HSC) play a central role in scar formation that leads to liver fibrosis. The molecular mechanisms underlying this process are not fully understood. Microarray and bioinformatics analyses have proven to be useful in identifying transcription factors that regulate cellular processes such as cell differentiation. Using oligonucleotide microarrays, we performed transcriptional analyses of activated human HSC cultured on Matrigel-coated tissue culture dishes. Examination of microarray data following Matrigel-induced deactivation of HSC revealed a significant down-regulation of myocardin, an important transcriptional regulator in smooth and cardiac muscle development. Thus, gene expression profiling as well as functional assays of activated HSC have provided the first evidence of the involvement of myocardin in HSC activation.

No MeSH data available.


Related in: MedlinePlus