Naslov
Morphological analysis of rat embryo differentiation in serum-free culture in vitro

Autori
Belovari, Tatjana ; Stević, Nataša ; Gajović, Srećko ; Kostović-Knežević, Ljiljana

Vrsta, podvrsta i kategorija rada
Radovi u zbornicima skupova, cjeloviti rad (in extenso), znanstveni

Izvornik
Proceedings 6th Multinational Congress on Microscopy-European Extension / Milat, Ognjen ; Ježek, Davor - Zagreb : Hrvatsko mikroskopijsko društvo, 2003, 314-15

Skup
6th Multinational Congress on Microscopy-European Extension

Mjesto i datum
Pula, Hrvatska, 01.06.2003. - 05.06.2003

Vrsta sudjelovanja
Poster

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
serum-free; differentiation; ultrastructure

Sažetak
An advantage of serum-free culture models in investigation of regulatory factors in embryo development is its precise formulation and avoiding of variability in serum composition (Barnes and Sato 1980). Modifications of in vitro conditions can promote or impede growth and differentiation of cultivated embryo. When gastrulating rat embryo is cultivated in serum-free medium only epidermis, cartilage, gut and respiratory epithelium differentiate regularly in contrast with the serum-supplemented medium, where also neuroblasts and myotubes develop (Škreb et al. 1993). Although light microscope analysis shows that observed tissues are terminally differentiated, imunohistochemical analysis suggests that undifferentiated cells exist in all tissues (Belovari et al. 2001). In order to determine the degree of tissue differentiation in cultivated rat embryos electron microscopy was used to analyse their ultrastructural characteristics. Gastrulating rat embryos (E 9.5) were cultivated for 14 days in chemically defined Eagle's Minimal Essential Medium (MEM) alone, and in MEM with human transferrin (50 μ g/ml), fixed afterwards with 4% glutaraldehide and 1% OsO4, embedded in Durcopan, sectioned by ultramicrotome and observed with TEM Zeiss 902A. Electron microscopy showed in epidermis cell junctions, intermediate filaments and keratinisation. Columnar epithelium with cilia contained axoneme, and glandular epithelium contained secretory granules. Neuroblasts had typical nuclei and cell processes which formed cell to cell contacts. Fibroblasts and chondroblasts had abundant rough endoplasmic reticulum and synthesised components of extracellular matrix (Fig. 1) suggesting that functional maturation occured. It can be concluded that analyzed tissues reached a high degree of differentiation in vitro. Therefore the applied in vitro system can be useful for evaluation of histodifferentiation during normal development, as well as to determine the pathological changes on embryonic cells caused by embryotoxic substances.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti, Kliničke medicinske znanosti

POVEZANOST RADA