Uncompromised Quality: The Blueprint for Successful Primary Cell Culture

1. Uncompromised Quality: The Blueprint for Successful Primary Cell Culture Primary cells are gold standard to study cellular physiology, biochemistry (e.g. metabolic studies, aging, signaling mechanism etc.), the effects of drugs and toxic compounds. Primary Cells are defined as cells directly isolated from tissue biopsies. Primary cells closely mimic the phenotypes of cells making the original organ i.e. in vivo conditions. Utility of primary cells are constrained by its limited life span. It is crucial to maintain high-quality of primary cells to get reliable and consistent results. The quality of primary cells is influenced by multiple factors, such as lab environment, equipment, procedures, personnel etc. Here we have discussed how these factors are crucial to produce high- quality cells: Laboratory Design and Environment: Good Laboratory Practice (GLP) requirements of primary cells are implemented on the design, construction, equipment, quality control and oversight approvals of cell culture laboratory. Primary Cell culture laboratory comprises of cell culture laboratory, quarantine laboratory, quality control laboratory, freezer room and storage area. Primary Cell Culture Laboratory: Primary cell lines are generated from tissue biopsies or other materials e.g. blood, liposuction fluid etc. in cell culture laboratory. Before bringing any

2. cell lines or tissue to this laboratory it should be screened for mycoplasma or any other contaminants. Cell culture laboratory can be further segregated for different activities such as Cell culture, Media Preparation, waste disposal etc. to prevent cross-contamination. Quarantine Laboratory: Any foreign materials entering in laboratory are referred as “Quarantine” materials until are tested free of contaminants such as bacteria, fungi and particularly mycoplasma. These foreign materials are stored in quarantine laboratory until tested for contaminants. Quality Control Laboratory: The quality control (QC) laboratory is equipped with all testing instruments required to evaluate quality standards of primary cells rigorously. Freezer Room: Cryopreservation techniques for long-term storage of primary cells are prerequisite for commercial uses. Establishment of efficient cryopreservation and storage techniques for primary cells is required. Primary Cell Laboratories requires multiple temperature freezers such as controlled rate freezers (-200C), low temperature freezers (-70 to -90 0C) and cry storage at -135 0 Backup generators and warning systems are required for these freezers. Storage Area: Chemicals and reagents used in cell culture laboratory should be clearly labelled and stored in designated place. Expired chemicals should be discarded immediately. Hazardous and non-compatible chemicals should be stored properly. All chemicals should be labelled with the date it was received and opened. FIGURE 1: (a) & (b) Change room for change street cloth to scrub; (c) Waste disposal via waste hatch; (d) Documentation area; (e) Lab Corridor; (f) Production personnel working in cleanroom. Image Resource: ISBN: 978-981-16-7589-8

3. Equipment and Facility Maintenance and Calibration Equipment plays a crucial role in primary cell culture laboratory. Equipment is required to produce consistent and good quality products. Hence, all equipment needs to undergo Installation Qualification (IQ), Operational Qualification (OQ), and Performance Qualification (PQ) to ensure suitability for its intended use. Facility should be validated for Heating, Ventilation and Air Conditioning (HVAC) system, cleaning, operator capacity etc. Equipment should be well monitored and calibrated regularly according to manufacturer’s instructions. Standard Operating Procedures (SOPs) All cell culture techniques, media preparation and equipment usage should be performed following stringent SOPs. All lab personnel should be trained and proficient in performing all procedures. Cell culture practices All primary cell culture procedure should be performed aseptically to prevent any contaminations. All reagents, equipment and consumables should be sterile. Cell culture media should be prepared and stored following manufacturer’s instructions and SOPs. pH and sterility of media should be checked before use. Cells should be handled gently to prevent mechanical stress. Cell morphology and growth should be monitored regularly. Quality Control Primary cells should be regularly monitored for microbial contaminants such as bacteria, fungi, and mycoplasma. Cell line should be verified and identified by STR profiling. Specific cell functionality should be evaluated regularly. Record Keeping and Documentation Comprehensive documentation of all cell culture activity should be recorded such as cell line origin, cell passaging, media change and other observed issues. Personal Hygiene and Safety Lab personnel should wear appropriate Personal Protective Equipment (PPE) such lab coat/cover all suit, gloves, lab shoes, head cap, face mask etc. Hand should be washed before and after cell culture work. Regular training should be provided to all lab personnel on biosafety and emergency procedures.

4. Consumables and Reagents Consumables and reagents should be sources from reputed manufacturers, and quality should be verified upon receipt. Reagents and consumables should be stored at appropriate conditions as per manufacturer’s instructions. Cell Banking and Cryopreservation Primary Cells should be cryopreserved following optimized SOPs, cell viability should be monitored regularly. Master bank and working bank should be established for cell lines. Compliance and Auditing Regular internal audits should be performed regularly for continuous improvement. Laboratory should be complied with relevant regulatory guidelines such as ISO, GLP (Good Laboratory Practices), GMP (Good Manufacturing Practices) etc. Information Technology Laboratory should be equipped with at least one computer system to maintain experimental documentation. FIGURE 2: (a–c) Building and equipment monitoring system; (d) CCTV security camera; (e) Equipment for environmental monitoring (air sampler and particle counter); (f) Equipment maintenance and service by authorised personnel; (g) Facility personnel cleaning. Image Resource: ISBN: 978-981-16-7589-8

5. By adhering to these quality aspects at Kosheeka, we have achieved highest standards of primary cell culture, minimised contamination risks and provide reliable and consistent results. Kosheeka is committed to provide high-quality primary cell culture Services by following stringent cell culture techniques, rigorous quality control testing, dedicated lab facility and scientific team. CONTACT US +91-9654321400 www.advancells.com Info@advancells.com