**1. Introduction**

Cell culture or Chicken embryo fibroblast cell culture is a fundamental laboratory technique that widely used in virology, vaccinology, molecular biology, microbiology as well as in biotechnology field. In this *in vitro* study controlled environment like media, pH, CO2 level, temperature, humidity, O2 flow mimics *in vivo* condition for cell to grow. Within last past decades a few species have been considered *in vitro*

utilizing cell culture technique exposing essential data regarding their biology. The chicken or avian embryo gives an amazing model for the study embryology, developmental biology and production of pharmaceutical proteins in transgenic chickens moreover take part in virology, immunization advancement as well as vaccinology research. Chicken embryo fibroblast cell culture has great impact on veterinary vaccine and biochemical production research. Besides, chicken stem cells have given a perfect opportunity for producing cell-based transgenic birds and a powerful source of cells for vaccine production for poultry and human viral diseases. Besides, they give knowledge into fundamental science namely drug sensitivity testing, cell tracking and cytotoxicity testing [1]. However, various important virus or viral diseases have never been explored at a cellular level so that cell culture technology is still in an early stage of its potential progress. In this document, it is summarized the basic steps engaged in the establishment of primary cell culture and concept of cell culture technique to serve as a practical guide for current and future researcher to leverage the power of cell cultures. This approach has the potential to produce valuable outcomes and suggestions regarding CEF cell culture, cell metabolism, adaptation to different stresses and challenges. Although most published papers discussed on CEF cell culture, BHK-21 and other cell lines, here focused on basic concept and necessary measurement for primary cell culture that inevitably helpful for novice as well as experienced researchers or laboratory personnel of this field [2]. Physiological and physiochemical condition: Nature and impact of pH level, temperature, concentration of O2/CO2, and osmotic pressure (culture media) that can be altered the result of the study as well as their effects on the cell culture [3]. Cell culture technology has spread productively within a century; a variety of culture media has been designed. This chapter goes through the brief concept, challenges and current issues of cell culture or chicken embryo fibroblast cell culture technique. Current article does not substance to be a complete guide but rather helps the researcher to plan and make consonant decision to their research experimental or work.
