This book covers cell culture technologies, providing an overview of isolating cells from their native tissue and cultivating them in a controlled, artificial laboratory environment. This process is a fundamental technique in red and green biotechnology and life science research. Essential considerations regarding cryopreservation, recovery, and mycoplasma contamination, along with how to prevent and treat them in primary cell cultures, are discussed. This book presents updated research findings on primary cell isolation from tissues, the generation of differentiated cells from pluripotent stem cells, their in vitro growth in 2-D or 3-D, and their analysis using modern technologies. Furthermore, the use of organoids in a complex tumour-on-a-chip model is discussed. In addition to the medical application of cell cultures, plant cell culture-specific points are also addressed. This book is a valuable resource for anyone studying or researching primary cell culture applications and techniques, offering invaluable insights into emerging trends and techniques.