Preface

Welcome to *Technologies in Cell Culture – A Journey from Basics to Advanced Applications*. In the ever-evolving landscape of biotechnology, cell culture is a cornerstone that enables innovations and amazing discoveries in fields ranging from basic cell biology to medicine, with current and upcoming research on biocomputing. This book is a comprehensive exploration of this vital discipline, offering a roadmap that navigates from fundamental principles to cutting-edge methodologies.

The goal of this book is to give a clear picture of the current state-of-the-art technological and application updates in cell culture. It is a useful handbook for students, researchers, and practitioners in the field. We hope that it will inspire future research ideas and directions.

In a time when scientific discoveries have the potential to both lessen human suffering and advance our knowledge of intricate biological systems, the chapters in this book act as guiding lights toward advancement. This book unravels the complexities of cell culture and provides both beginners and experienced researchers with a clear understanding of the underlying principles, techniques, and applications. Each chapter marks a significant step in the quest for knowledge and understanding, ranging from the fundamentals of cell culture to the cutting edge of sophisticated applications.

This book includes eleven chapters organized into six sections based on topics related to cell culture technological advancements and applications.

Section 1 consists of four chapters that narrate methods of two-dimensional (2D) and three-dimensional (3D) cell cultures. Chapter 1 describes valvular interstitial cellular (VIC) physiology, as well as the isolation and culture of these types of cells. In our aging world, valve pathology is becoming more common than ever, and there is very little information available regarding VIC pathophysiology. VIC stable cell populations in the lab are necessary for the scientific investigation of valve disease. These populations are frequently isolated from tissue from pigs and humans. For experimental integrity to be maintained, it is crucial to comprehend the many VIC phenotypes as well as the molecular cues that control the transition between phenotypes. Thus, this chapter presents an outline of VIC physiology and a triedand-true technique for isolating and cultivating these cells. Chapter 2 provides a comprehensive review of culturing techniques for human pluripotent stem cells (hPSCs), covering 2D and 3D adherent culture, suspension culture, and the utilization of hydrogel scaffolds in 3D hPSC culture. To meet the demands of cutting-edge biomedical applications, the chapter also investigates the use of sophisticated 3D cell manufacturing techniques to simplify the creation of vast quantities of high-quality hPSCs. By covering these subjects, this chapter provides a thorough summary of various culturing techniques and the many uses they have in hPSC research, including basic research and cutting-edge biomedical studies.

The study of interactions between tumor microenvironment cells, including adipocytes, endothelial cells, fibroblasts, macrophages, and cancer cells, is commonly conducted by *in vitro* coculture techniques. Indirect and direct methods are the two primary categories into which coculture techniques are divided. Chapter 3 discusses the general techniques for coculture research, focusing on the outcomes of coculturing cancer cells and adipocytes. Chapter 4 describes the formation of spontaneously contracting heart primary cultures obtained from fish embryos and larvae. These cultures are useful models for several purposes, including studying heart-related viral infections and conducting research in biomedicine, pharmacology, and cardiology. Chapter 4 also presents a summary of the findings regarding the *in vitro* production of 3D heart-like cell aggregates from fully digested fish embryos and larvae.

To evaluate and validate the qualities of plant-based natural substances for use as possible therapies, Chapter 5 in Section 2 addresses the latest technological advancements in cell-free and cell-based approaches for determining properties like induction of oxidative DNA damage, inflammatory factors, apoptotic/anti-apoptotic factors, and so on.

In Section 3, Chapters 6 and 7 discuss the application of cell culture in enhancing drug discovery. Chapter 6 offers a thorough analysis of the traits and attributes of *in vitro* techniques that bolster the efficacy of two compounds. Cytotoxicity assays in cell lines and primary cell cultures of pancreatic and breast cancer grown in 2D and 3D conditions are used to determine the efficacy of both molecules, specifically against mutant forms of K-Ras as antitumor agents. Recent technologies for cultivating cells have recently been developed to mimic the biophysical characteristics of tumors. With the use of these new technologies, it is possible to create 3D tumor-like structures, or spheroids, in which cancer cells connect in a manner akin to that seen in actual tumors. As a result of these developments in cell culture technology, cell-based assays must be modified and improved. Chapter 7 focuses on certain problems encountered and the solutions found during the assessment of the effects of two anticancer medications (temozolomide and vemurafenib) on spheroids-grown melanoma cell lines.

In Section 3, Chapter 8 discusses autologous adipose tissue stem cell-derived terminally differentiated retinal pigment epithelium monolayer cell development on a biocompatible scaffold in the restoration of vision in age-related macular degeneration patients.

Section 5 includes Chapters 9 and 10, which discuss the application of cell culture in bioprocessing. Chapter 9 highlights the recent advancements in process analytical technology-based tool development, specifically for biopharmaceutical applications, along with the techniques inherited from the traditional pharmaceutical sector to monitor and control critical process parameters and critical quality attributes. Considering current and upcoming trends to address bottlenecks and problems, Chapter 10 discusses techniques for manufacturing lentiviral vectors in HEK293 cells grown in serum-free media and the concepts of optimizing along with scaling-up of bioprocesses.

Finally, Chapter 11 in Section 6 discusses the most recent development and application of cell culture technology, the intersection of biology and machine learning and organoid intelligence (OI), which can reshape our understanding of life and health. The goal of the OI concept is to interface brain organoids with computer technology

in order to utilize their inherent biological capabilities for biocomputing and synthetic intelligence memorization.

We hope this book will suffice its purpose of disseminating knowledge to the students and scientific community.

We would like to acknowledge the authors and reviewers for their excellent contributions. We also wish to thank the staff at IntechOpen for their enormous support.

## **Dr. Soumya Basu**

Associate Professor, Cancer and Translational Research Centre, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India

## **Dr. Amit Ranjan**

Assistant Professor, Cancer and Translational Research Centre, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India

## **Dr. Subhayan Sur**

DBT-Ramalingaswami Re-Entry Fellow, Assistant Professor, Cancer and Translational Research Centre, Dr. D.Y. Patil Biotechnology and Bioinformatics Institute, Dr. D. Y. Patil Vidyapeeth, Pune, Maharashtra, India
