Reviewer's report 
 Long-term culture of cholangiocytes from liver fibro-granulomatous lesions Title: 
 2  20 January 2006 Version: Date: 
 Eugenio Gaudio Reviewer: 
Reviewer's report: 
General 
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Major Compulsory Revisions (that the author must respond to before a decision on publication can be reached) 
1. The authors should improve the description of optical microscopic methods. 
2. Did the Authors use a method to block unspecific binding of the antibodie in the 
immunofluorescence stainings? and a method to permeabilize the cells (for protein expressed 
intracellularly)? 
3. In the immunofluorescence methods, the authors should indicate the description of primary and 
secondary antibodies (species reactivity, sources and, if possible, the company that provided the 
primary antibody), the dilution used and the time of incubation with the secondary antibodies. 
4. Figure 3A: the H&E staining should be changed with a better section (the nuclei are not well 
displayed) or/and with a section with a Masson’s trichrome stain. 
5. In the discussion the authors do not explain why they have been not able to isolate cholangiocytes 
from dissociated chronic phase granulomas or dissociated liver fragments. This aspect should be 
better elucidated in the discussion. 
6. In the discussion some sentences are not supported by references. 
7. The cell line should be tested for epithelial markers (CK-19 at least) at least after 16 passages 
and at one year and eight months of culture 
------------------------------------------------------------------------------- 
Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the 
author can be trusted to correct) 
1. Legend figure 3, line13: microvilli instead of villi. 
2. Legend figure 5, line 2: the label “B.” misses. 
3. Figure 1. The chronic phase is described as 45 days after the infection instead of 90 days. 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Unable to decide on acceptance or rejection until the authors have responded to the What next?: 
major compulsory revisions 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
Reviewer's report 
 Long-term culture of cholangiocytes from liver fibro-granulomatous lesions Title: 
 2  20 January 2006 Version: Date: 
 Eugenio Gaudio Reviewer: 
Reviewer's report: 
General 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can be reached) 
1. The authors should improve the description of optical microscopic methods. 
2. Did the Authors use a method to block unspecific binding of the antibodie in the 
immunofluorescence stainings? and a method to permeabilize the cells (for protein expressed 
intracellularly)? 
3. In the immunofluorescence methods, the authors should indicate the description of primary and 
secondary antibodies (species reactivity, sources and, if possible, the company that provided the 
primary antibody), the dilution used and the time of incubation with the secondary antibodies. 
4. Figure 3A: the H&E staining should be changed with a better section (the nuclei are not well 
displayed) or/and with a section with a Masson’s trichrome stain. 
5. In the discussion the authors do not explain why they have been not able to isolate cholangiocytes 
from dissociated chronic phase granulomas or dissociated liver fragments. This aspect should be 
better elucidated in the discussion. 
6. In the discussion some sentences are not supported by references. 
7. The cell line should be tested for epithelial markers (CK-19 at least) at least after 16 passages 
and at one year and eight months of culture 
------------------------------------------------------------------------------- 
Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the 
author can be trusted to correct) 
1. Legend figure 3, line13: microvilli instead of villi. 
2. Legend figure 5, line 2: the label “B.” misses. 
3. Figure 1. The chronic phase is described as 45 days after the infection instead of 90 days. 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Unable to decide on acceptance or rejection until the authors have responded to the What next?: 
major compulsory revisions 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
Please review this paper.
user
Reviewer's report 
 Long-term culture of cholangiocytes from liver fibro-granulomatous lesions Title: 
 2  20 January 2006 Version: Date: 
 Eugenio Gaudio Reviewer: 
Reviewer's report: 
General 
------------------------------------------------------------------------------- 
Major Compulsory Revisions (that the author must respond to before a decision on publication can be reached) 
1. The authors should improve the description of optical microscopic methods. 
2. Did the Authors use a method to block unspecific binding of the antibodie in the 
immunofluorescence stainings? and a method to permeabilize the cells (for protein expressed 
intracellularly)? 
3. In the immunofluorescence methods, the authors should indicate the description of primary and 
secondary antibodies (species reactivity, sources and, if possible, the company that provided the 
primary antibody), the dilution used and the time of incubation with the secondary antibodies. 
4. Figure 3A: the H&E staining should be changed with a better section (the nuclei are not well 
displayed) or/and with a section with a Masson’s trichrome stain. 
5. In the discussion the authors do not explain why they have been not able to isolate cholangiocytes 
from dissociated chronic phase granulomas or dissociated liver fragments. This aspect should be 
better elucidated in the discussion. 
6. In the discussion some sentences are not supported by references. 
7. The cell line should be tested for epithelial markers (CK-19 at least) at least after 16 passages 
and at one year and eight months of culture 
------------------------------------------------------------------------------- 
Minor Essential Revisions (such as missing labels on figures, or the wrong use of a term, which the 
author can be trusted to correct) 
1. Legend figure 3, line13: microvilli instead of villi. 
2. Legend figure 5, line 2: the label “B.” misses. 
3. Figure 1. The chronic phase is described as 45 days after the infection instead of 90 days. 
------------------------------------------------------------------------------- 
Discretionary Revisions (which the author can choose to ignore) 
 Unable to decide on acceptance or rejection until the authors have responded to the What next?: 
major compulsory revisions 
 An article whose findings are important to those with closely related research Level of interest: 
interests 
 Acceptable Quality of written English: 
assistant