Design-based stereological estimation of hepatocyte number, by combining the smooth optical fractionator and immunocytochemistry with anticarcino embryonic antigen polyclonal antibodies

• Type: Article in International Scientific Journal
• Date: 2006

Authors

marcos, r (Author) Other; monteiro, raf (Author) ICBAS; rocha, e (Author) ICBAS

Journal

Title: Liver International

Vol. 26

Pages: 116-124

ISSN: 1478-3223

Publisher: Wiley-Blackwell

Indexing

ISI Web of Knowledge - 36 Citations

Scopus - 39 Citations

Scientific classification

FOS: Medical and Health sciences > Clinical medicine

Other information

Authenticus ID: P-004-NHJ

DOI: 10.1111/j.1478-3231.2005.01201.x

Abstract (EN): Backgound/Aims: Hepatocytes (HEP) have been the major target for structural quantification in the liver, but an estimation of their total number (N), their percentage in relation to the global number of liver cells and the evaluation of the percentage of binucleated hepatocytes (BnHEPs) have never been performed with modern design-based stereological techniques. The establishment of sound technical guidelines and baseline quantitative data in non-pathological conditions are relevant to properly evaluate HEP hyperplasia and BnHEP responses. Methods: In this study, we combined immunocytochemistry with sound design-based stereology for estimating the N of HEP and the N of non-hepatocytic cells (NHCs). For obtaining systematic uniform random sections (30 mu m thick), a smooth fractionator sampling scheme was applied to the liver of five mate Wistar rats (3 month old). Those sections were immunostained with polyclonal antibodies against carcinoembryonic antigen. Because biliary canaliculi were then marked, an unequivocal counting of mononucleated hepatocytes (MnHEP) and BnHEP was allowed. Results: The N of HEP was estimated to be 1.93x10(9), with a coefficient of error (CE) of 0.02, corresponding to 129x10(6) HEP/g of liver. BnHEP represented 26% of total HEP number. The N of NHC was estimated as 1.31x10(9) (CE=0.02). Conclusion: The strategy here presented provides a reliable method for accessing the N of HEP (distinguishing MnHEP from BnHEP) in situations in which these parameters are relevant, namely for evaluating the magnitude of an hyperplastic liver response from its very early onset.

Language: English

Type (Professor's evaluation): Scientific

No. of pages: 9