Establishment Of Culture System Of Roe Deer Fibroblast In Vitro And Analysis Of Biological Characteristics

The roe deer(Capreolus gray)is divided into two species,which contain Eastern roe deer(Siberian roe deer)and Western roe deer(European roe deer),the Siberian roe deer are distributing in Inner Mongolia of China,and the population of roe deer has been significantly reduced in this region.This study aims to establish cultivation system of roe deer fibroblasts in vitro and analyze its biological characteristics,try to protect the roe deer species resources.Futhermore,to build cervidae animal phylogenetic tree to explore relationships of cervid family.In this study,fibroblast lineage was established from male and female roe deer by tissue adherent culture method,and the established cell line were consistent with the morphology and growth characteristics of fibroblasts.Next,we analyzed the biological characteristics of fibroblasts from three tissue sources of roe deer,which including measured the attachment rate of the three kinds of fibroblasts,the cell survival rate before and after cryopreservation,and ploted the cell growth curve of the trachea and ear fibroblasts showed typical S-type of cell growthing.The H&E staining of fibroblasts from three type of tissues showed that the cells grew in a swirl shape,and the cell morphology was fusiformis,triangular or irregular,which was the typical morphology of fibroblasts.The karyotype and G-band analysis results of male and female roe deer showed that the number of chromosomes was 2n=70(include XY/XX),and also showing genetic stability was confirmed.The efficiency of transfection of green fluorescent protein granules(p EGFP-C1)was more than 20% at12 h.After transfection of red fluorescent protein granules(H2B-CAG-td Tomato),trachea and ear fibroblast cell lines expression stable red fiuorescence by screening.In this study,phylogenetic trees of 21 species of cervidae were constructed based on the analysis of 5 combined mitochondrial genes,which can visually display the phylogenetic relationships of deer family,Siberian roe deer are closely related with reindeer.The roe deer genetic resources and information obtained in this study were finally recorded into the Bank of Inner Mongolia Plateau Animal Genetic Resources Information Database,which realized the protected wild animal genetic resources and provided useful data and material to support further research in this species.