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In the present study, human neonatal fibroblasts were isolated from a two-month-old human male. The purpose of the present investigation was the analysis of the morphology (light and transmission electron microscopy), karyotype and growth characteristics of the human neonatal fibroblast cell culture B-HNF-1. Moreover, STR typing and mitochondrial DNA amplification and sequencing was also performed. Analysis of chromosomes count showed that B-HNF-1 cell culture is diploid and has normal male karyotype 46, XY, which was stable during cultivation. The transmission electron microscopy demonstrated the ultra-structure of the B-HNF-1 cells; they have typical morphological features of proteosynthesis-active cells. Large number of fibroblasts bearing different shapes and surface characteristics adhered to the substrate with microvilli and filopodia. Our in vitro expanded fibroblasts have a large and irregular nucleus with prevalence of euchromatin. One to three nucleoli are present in each nucleus. The cytoplasm contains a richly developed rough endoplasmic reticulum and prominent Golgi apparatus cisterns. The result of the fragment analysis is a DNA profile defined as multiplex of STR markers and sex determining system. Sequencing analysis of hypervariable segments of control region of mitochondrial DNA showed haplotype that belongs to haplogroup W. In this study, we complexly characterized a new cell culture of human neonatal fibroblasts B-HNF-1 from different points of view. This culture should be used for further biomedical experiments.

Biological and morphological characterization of human neonatal fibroblast cell culture B-HNF-1