Reviewer's report
Title:Mouse model of carbon tetrachloride induced liver fibrosis:
Histopathological changes and expression of CD133 and epidermal growth factor
Version: 1Date:16 December 2009
Reviewer: Frank Tacke
Reviewer's report:
Fujii et al describe experiments in which mice were treated with CCl4 per
gavage, resulting in liver fibrosis and tumor formation. They also found
upreguated gene expression of EGF, TGF, HSC markers and CD133 and
conclude that CD133 might be involved in fibrosis progression or tumor
formation.
Major Compulsory Revisions
1. It is very hard to believe that treatment with CCl4 per gavage leads to liver
fibrosis and HCC development. CCl4 has been extensively used in the literature
(typically i.p. injections for 6-8 weeks), but usually does not result in HCC. If this
should be true, the authors need to provide compelling experimental evidence.
To prove fibrosis formation: hydroxyproline assay, collagen or a-SMA Western
Blot, collagen immunohistochemistry. For tumor formation: clonal analysis,
macroscopic picture, gene profile from tumorous vs non-tumorous tissue.
2. The paper is solely descriptive, and the up-regulation of HSC-related genes in
a fibrosis model is not surprising. If the authors believe that they have HCC
development, they should differentiate gene expression profiles at time-point III
for tumorous vs non-tumorous tissue.
3. It is completely unclear if the upregulation of CD133 has anything to do with
HSC activation or proliferation. Therefore, clear immunohistochemical stainings
are needed to assess CD133 in liver tissue and assess co-staining with activated
HSC (e.g., by a-SMA staining). Many hematopoietic cells can express CD133 as
well, this should be tested. Furthermore, it would be helpful to isolate primary
cells from the treated livers, i.e. hepatocytes, HSC, endothelial cells and Kupffer
cells, to assess which cell type expresses CD133. The use of cell lines is not
helpful in this respect.
Level of interest: An article of limited interest
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
Reviewer's report
Title:Mouse model of carbon tetrachloride induced liver fibrosis:
Histopathological changes and expression of CD133 and epidermal growth factor
Version: 1Date:16 December 2009
Reviewer: Frank Tacke
Reviewer's report:
Fujii et al describe experiments in which mice were treated with CCl4 per
gavage, resulting in liver fibrosis and tumor formation. They also found
upreguated gene expression of EGF, TGF, HSC markers and CD133 and
conclude that CD133 might be involved in fibrosis progression or tumor
formation.
Major Compulsory Revisions
1. It is very hard to believe that treatment with CCl4 per gavage leads to liver
fibrosis and HCC development. CCl4 has been extensively used in the literature
(typically i.p. injections for 6-8 weeks), but usually does not result in HCC. If this
should be true, the authors need to provide compelling experimental evidence.
To prove fibrosis formation: hydroxyproline assay, collagen or a-SMA Western
Blot, collagen immunohistochemistry. For tumor formation: clonal analysis,
macroscopic picture, gene profile from tumorous vs non-tumorous tissue.
2. The paper is solely descriptive, and the up-regulation of HSC-related genes in
a fibrosis model is not surprising. If the authors believe that they have HCC
development, they should differentiate gene expression profiles at time-point III
for tumorous vs non-tumorous tissue.
3. It is completely unclear if the upregulation of CD133 has anything to do with
HSC activation or proliferation. Therefore, clear immunohistochemical stainings
are needed to assess CD133 in liver tissue and assess co-staining with activated
HSC (e.g., by a-SMA staining). Many hematopoietic cells can express CD133 as
well, this should be tested. Furthermore, it would be helpful to isolate primary
cells from the treated livers, i.e. hepatocytes, HSC, endothelial cells and Kupffer
cells, to assess which cell type expresses CD133. The use of cell lines is not
helpful in this respect.
Level of interest: An article of limited interest
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
Please review this paper.
user
Reviewer's report
Title:Mouse model of carbon tetrachloride induced liver fibrosis:
Histopathological changes and expression of CD133 and epidermal growth factor
Version: 1Date:16 December 2009
Reviewer: Frank Tacke
Reviewer's report:
Fujii et al describe experiments in which mice were treated with CCl4 per
gavage, resulting in liver fibrosis and tumor formation. They also found
upreguated gene expression of EGF, TGF, HSC markers and CD133 and
conclude that CD133 might be involved in fibrosis progression or tumor
formation.
Major Compulsory Revisions
1. It is very hard to believe that treatment with CCl4 per gavage leads to liver
fibrosis and HCC development. CCl4 has been extensively used in the literature
(typically i.p. injections for 6-8 weeks), but usually does not result in HCC. If this
should be true, the authors need to provide compelling experimental evidence.
To prove fibrosis formation: hydroxyproline assay, collagen or a-SMA Western
Blot, collagen immunohistochemistry. For tumor formation: clonal analysis,
macroscopic picture, gene profile from tumorous vs non-tumorous tissue.
2. The paper is solely descriptive, and the up-regulation of HSC-related genes in
a fibrosis model is not surprising. If the authors believe that they have HCC
development, they should differentiate gene expression profiles at time-point III
for tumorous vs non-tumorous tissue.
3. It is completely unclear if the upregulation of CD133 has anything to do with
HSC activation or proliferation. Therefore, clear immunohistochemical stainings
are needed to assess CD133 in liver tissue and assess co-staining with activated
HSC (e.g., by a-SMA staining). Many hematopoietic cells can express CD133 as
well, this should be tested. Furthermore, it would be helpful to isolate primary
cells from the treated livers, i.e. hepatocytes, HSC, endothelial cells and Kupffer
cells, to assess which cell type expresses CD133. The use of cell lines is not
helpful in this respect.
Level of interest: An article of limited interest
Quality of written English: Acceptable
Statistical review: No, the manuscript does not need to be seen by a
statistician.
Declaration of competing interests:
I declare that I have no competing interests
assistant