Reviewer's report
Title:Co-culture of primary human tumor hepatocytes from patients with
hepatocellular carcinoma with autologous peripheral blood mononuclear cells:
Study of their in vitro immunological interactions.
Version: 1Date:3 August 2012
Reviewer: Frank Tacke
Reviewer's report:
This manuscript describes the culture of primary parenchymal cells from normal
human liver and HCC specimens. Furthermore, the authors conducted
co-cultures with autologous PBMCs.
1. Although I acknowledge the technical value of the culture of primary
parenchymal human liver cells, I believe the work is preliminary at present. The
FACS plots are not convincing at all, and the MHC-II expression of the
“hepatocytes” indicates that there might be “contamination” by other cells. The
authors only mention negative staining for CD14 or CD3, but this should be
shown. The quality of the presented FACS data is rather poor (likely due to the
low numbers of cells retrieved from human tissue), but this makes the data
difficult to interpret. I think it is important to show that (a) no cell doublets were
measured (FSC-W), and (b) all “hepatocytes” stain negative for CD45
(leukocytes), endothelial and HSC markers. Furthermore, immunohistochemistry
for typical KC and HSC markers (e.g., CD68, decorin) should be performed.
2. The co-culture experiments are, to my opinion, difficult to interpret. The
co-staining of CD8 and HLA-DR is not convincing, because the compensation
appears not appropriate. How should CD8+ cells and hepatocytes/HCC cells
“communicate”? Why do both populations upregulate MHC-II? I strongly doubt
that these observations are truly relevant. More functional assays would be
needed to confirm that these observations are not artefacts, but related to
pathogenically relevant mechanisms.
3. Co-cultures of hepatocytes / tumor cells with defined immune cell
subpopulations (e.g., isolated by MACS) are needed to identify and delineate the
interactions.
Level of interest: An article whose findings are important to those with closely
related research interests
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:Co-culture of primary human tumor hepatocytes from patients with
hepatocellular carcinoma with autologous peripheral blood mononuclear cells:
Study of their in vitro immunological interactions.
Version: 1Date:3 August 2012
Reviewer: Frank Tacke
Reviewer's report:
This manuscript describes the culture of primary parenchymal cells from normal
human liver and HCC specimens. Furthermore, the authors conducted
co-cultures with autologous PBMCs.
1. Although I acknowledge the technical value of the culture of primary
parenchymal human liver cells, I believe the work is preliminary at present. The
FACS plots are not convincing at all, and the MHC-II expression of the
“hepatocytes” indicates that there might be “contamination” by other cells. The
authors only mention negative staining for CD14 or CD3, but this should be
shown. The quality of the presented FACS data is rather poor (likely due to the
low numbers of cells retrieved from human tissue), but this makes the data
difficult to interpret. I think it is important to show that (a) no cell doublets were
measured (FSC-W), and (b) all “hepatocytes” stain negative for CD45
(leukocytes), endothelial and HSC markers. Furthermore, immunohistochemistry
for typical KC and HSC markers (e.g., CD68, decorin) should be performed.
2. The co-culture experiments are, to my opinion, difficult to interpret. The
co-staining of CD8 and HLA-DR is not convincing, because the compensation
appears not appropriate. How should CD8+ cells and hepatocytes/HCC cells
“communicate”? Why do both populations upregulate MHC-II? I strongly doubt
that these observations are truly relevant. More functional assays would be
needed to confirm that these observations are not artefacts, but related to
pathogenically relevant mechanisms.
3. Co-cultures of hepatocytes / tumor cells with defined immune cell
subpopulations (e.g., isolated by MACS) are needed to identify and delineate the
interactions.
Level of interest: An article whose findings are important to those with closely
related research interests
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:Co-culture of primary human tumor hepatocytes from patients with
hepatocellular carcinoma with autologous peripheral blood mononuclear cells:
Study of their in vitro immunological interactions.
Version: 1Date:3 August 2012
Reviewer: Frank Tacke
Reviewer's report:
This manuscript describes the culture of primary parenchymal cells from normal
human liver and HCC specimens. Furthermore, the authors conducted
co-cultures with autologous PBMCs.
1. Although I acknowledge the technical value of the culture of primary
parenchymal human liver cells, I believe the work is preliminary at present. The
FACS plots are not convincing at all, and the MHC-II expression of the
“hepatocytes” indicates that there might be “contamination” by other cells. The
authors only mention negative staining for CD14 or CD3, but this should be
shown. The quality of the presented FACS data is rather poor (likely due to the
low numbers of cells retrieved from human tissue), but this makes the data
difficult to interpret. I think it is important to show that (a) no cell doublets were
measured (FSC-W), and (b) all “hepatocytes” stain negative for CD45
(leukocytes), endothelial and HSC markers. Furthermore, immunohistochemistry
for typical KC and HSC markers (e.g., CD68, decorin) should be performed.
2. The co-culture experiments are, to my opinion, difficult to interpret. The
co-staining of CD8 and HLA-DR is not convincing, because the compensation
appears not appropriate. How should CD8+ cells and hepatocytes/HCC cells
“communicate”? Why do both populations upregulate MHC-II? I strongly doubt
that these observations are truly relevant. More functional assays would be
needed to confirm that these observations are not artefacts, but related to
pathogenically relevant mechanisms.
3. Co-cultures of hepatocytes / tumor cells with defined immune cell
subpopulations (e.g., isolated by MACS) are needed to identify and delineate the
interactions.
Level of interest: An article whose findings are important to those with closely
related research interests
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.
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