To master the basic operation process of primary culture and subculture of mammalian cells, lay a foundation for the application of bioengineering in medicine.
Contains everything about cell culture and cell culture laboratory. The data has been collected from various sources and piled up to make this presentation.
India Clinical Trials Market: Industry Size and Growth Trends [2030] Analyzed...Kumar Satyam
According to TechSci Research report, "India Clinical Trials Market- By Region, Competition, Forecast & Opportunities, 2030F," the India Clinical Trials Market was valued at USD 2.05 billion in 2024 and is projected to grow at a compound annual growth rate (CAGR) of 8.64% through 2030. The market is driven by a variety of factors, making India an attractive destination for pharmaceutical companies and researchers. India's vast and diverse patient population, cost-effective operational environment, and a large pool of skilled medical professionals contribute significantly to the market's growth. Additionally, increasing government support in streamlining regulations and the growing prevalence of lifestyle diseases further propel the clinical trials market.
Growing Prevalence of Lifestyle Diseases
The rising incidence of lifestyle diseases such as diabetes, cardiovascular diseases, and cancer is a major trend driving the clinical trials market in India. These conditions necessitate the development and testing of new treatment methods, creating a robust demand for clinical trials. The increasing burden of these diseases highlights the need for innovative therapies and underscores the importance of India as a key player in global clinical research.
To master the basic operation process of primary culture and subculture of mammalian cells, lay a foundation for the application of bioengineering in medicine.
Contains everything about cell culture and cell culture laboratory. The data has been collected from various sources and piled up to make this presentation.
India Clinical Trials Market: Industry Size and Growth Trends [2030] Analyzed...Kumar Satyam
According to TechSci Research report, "India Clinical Trials Market- By Region, Competition, Forecast & Opportunities, 2030F," the India Clinical Trials Market was valued at USD 2.05 billion in 2024 and is projected to grow at a compound annual growth rate (CAGR) of 8.64% through 2030. The market is driven by a variety of factors, making India an attractive destination for pharmaceutical companies and researchers. India's vast and diverse patient population, cost-effective operational environment, and a large pool of skilled medical professionals contribute significantly to the market's growth. Additionally, increasing government support in streamlining regulations and the growing prevalence of lifestyle diseases further propel the clinical trials market.
Growing Prevalence of Lifestyle Diseases
The rising incidence of lifestyle diseases such as diabetes, cardiovascular diseases, and cancer is a major trend driving the clinical trials market in India. These conditions necessitate the development and testing of new treatment methods, creating a robust demand for clinical trials. The increasing burden of these diseases highlights the need for innovative therapies and underscores the importance of India as a key player in global clinical research.
Explore our infographic on 'Essential Metrics for Palliative Care Management' which highlights key performance indicators crucial for enhancing the quality and efficiency of palliative care services.
This visual guide breaks down important metrics across four categories: Patient-Centered Metrics, Care Efficiency Metrics, Quality of Life Metrics, and Staff Metrics. Each section is designed to help healthcare professionals monitor and improve care delivery for patients facing serious illnesses. Understand how to implement these metrics in your palliative care practices for better outcomes and higher satisfaction levels.
ICH Guidelines for Pharmacovigilance.pdfNEHA GUPTA
The "ICH Guidelines for Pharmacovigilance" PDF provides a comprehensive overview of the International Council for Harmonisation of Technical Requirements for Pharmaceuticals for Human Use (ICH) guidelines related to pharmacovigilance. These guidelines aim to ensure that drugs are safe and effective for patients by monitoring and assessing adverse effects, ensuring proper reporting systems, and improving risk management practices. The document is essential for professionals in the pharmaceutical industry, regulatory authorities, and healthcare providers, offering detailed procedures and standards for pharmacovigilance activities to enhance drug safety and protect public health.
Leading the Way in Nephrology: Dr. David Greene's Work with Stem Cells for Ki...Dr. David Greene Arizona
As we watch Dr. Greene's continued efforts and research in Arizona, it's clear that stem cell therapy holds a promising key to unlocking new doors in the treatment of kidney disease. With each study and trial, we step closer to a world where kidney disease is no longer a life sentence but a treatable condition, thanks to pioneers like Dr. David Greene.
One of the most developed cities of India, the city of Chennai is the capital of Tamilnadu and many people from different parts of India come here to earn their bread and butter. Being a metropolitan, the city is filled with towering building and beaches but the sad part as with almost every Indian city
Antibiotic Stewardship by Anushri Srivastava.pptxAnushriSrivastav
Stewardship is the act of taking good care of something.
Antimicrobial stewardship is a coordinated program that promotes the appropriate use of antimicrobials (including antibiotics), improves patient outcomes, reduces microbial resistance, and decreases the spread of infections caused by multidrug-resistant organisms.
WHO launched the Global Antimicrobial Resistance and Use Surveillance System (GLASS) in 2015 to fill knowledge gaps and inform strategies at all levels.
ACCORDING TO apic.org,
Antimicrobial stewardship is a coordinated program that promotes the appropriate use of antimicrobials (including antibiotics), improves patient outcomes, reduces microbial resistance, and decreases the spread of infections caused by multidrug-resistant organisms.
ACCORDING TO pewtrusts.org,
Antibiotic stewardship refers to efforts in doctors’ offices, hospitals, long term care facilities, and other health care settings to ensure that antibiotics are used only when necessary and appropriate
According to WHO,
Antimicrobial stewardship is a systematic approach to educate and support health care professionals to follow evidence-based guidelines for prescribing and administering antimicrobials
In 1996, John McGowan and Dale Gerding first applied the term antimicrobial stewardship, where they suggested a causal association between antimicrobial agent use and resistance. They also focused on the urgency of large-scale controlled trials of antimicrobial-use regulation employing sophisticated epidemiologic methods, molecular typing, and precise resistance mechanism analysis.
Antimicrobial Stewardship(AMS) refers to the optimal selection, dosing, and duration of antimicrobial treatment resulting in the best clinical outcome with minimal side effects to the patients and minimal impact on subsequent resistance.
According to the 2019 report, in the US, more than 2.8 million antibiotic-resistant infections occur each year, and more than 35000 people die. In addition to this, it also mentioned that 223,900 cases of Clostridoides difficile occurred in 2017, of which 12800 people died. The report did not include viruses or parasites
VISION
Being proactive
Supporting optimal animal and human health
Exploring ways to reduce overall use of antimicrobials
Using the drugs that prevent and treat disease by killing microscopic organisms in a responsible way
GOAL
to prevent the generation and spread of antimicrobial resistance (AMR). Doing so will preserve the effectiveness of these drugs in animals and humans for years to come.
being to preserve human and animal health and the effectiveness of antimicrobial medications.
to implement a multidisciplinary approach in assembling a stewardship team to include an infectious disease physician, a clinical pharmacist with infectious diseases training, infection preventionist, and a close collaboration with the staff in the clinical microbiology laboratory
to prevent antimicrobial overuse, misuse and abuse.
to minimize the developme
Deep Leg Vein Thrombosis (DVT): Meaning, Causes, Symptoms, Treatment, and Mor...The Lifesciences Magazine
Deep Leg Vein Thrombosis occurs when a blood clot forms in one or more of the deep veins in the legs. These clots can impede blood flow, leading to severe complications.
CHAPTER 1 SEMESTER V PREVENTIVE-PEDIATRICS.pdfSachin Sharma
This content provides an overview of preventive pediatrics. It defines preventive pediatrics as preventing disease and promoting children's physical, mental, and social well-being to achieve positive health. It discusses antenatal, postnatal, and social preventive pediatrics. It also covers various child health programs like immunization, breastfeeding, ICDS, and the roles of organizations like WHO, UNICEF, and nurses in preventive pediatrics.
Health Education on prevention of hypertensionRadhika kulvi
Hypertension is a chronic condition of concern due to its role in the causation of coronary heart diseases. Hypertension is a worldwide epidemic and important risk factor for coronary artery disease, stroke and renal diseases. Blood pressure is the force exerted by the blood against the walls of the blood vessels and is sufficient to maintain tissue perfusion during activity and rest. Hypertension is sustained elevation of BP. In adults, HTN exists when systolic blood pressure is equal to or greater than 140mmHg or diastolic BP is equal to or greater than 90mmHg. The
2. What is cell culture ?
• Cell culture is a technique which involves isolation of cells from
animal/plant body i.e. from their natural environment (in vivo) and
practicing to grow isolated cells in cell specific media in plastic flask
or petri dish in a controlled environmental artificial condition (in
vitro).
• Cell culture means to keep cells alive and grow in an in vitro
condition in a nutritive media which are widely used for research and
diagnosis of different pathogens and to understand the function and
mechanism of operation of many cells.
3. Types of cell culture
•Primary culture
Primary culture is the cell culture system that is formed by culture cells
directly obtained from tissue and proliferate them until they occupy all of the
available substrate (i.e. reach Confluence).
For primary culture, the first step is to isolate the tissue from organ and
splitting of cells from tissues. This can be achieved by trypsin treatment.
• Then grow cells in freshly prepared cell specific medium and incubate the
flask containing cells in incubator providing suitable environmental
condition for the growth of cells.
• After the first subculture, the primary culture is known as a cell
line or subclone.
4. •Secondary culture
• When cells from primary culture on confluency are isolated and
cultured in new media, it is called as secondary culture.
• Secondary culture is also known as subculture or splitting of cells.
• Subculture allows fresh nutrients and more space for the expansion
of the cells.
• Cells from primary culture are splits by trypsin/EDTA treatment.
• Trypsin which is a serine protease digest the extracellular protein or
matrix protein so that cells get free.
• EDTA which is a chelating agent chelates calcium ion because calcium
helps in cell adhesion.
5. Finite Vs Continuous Cell Line
• Normally, cells divide to a limited times, after that loose their proliferation
ability (senescence) these cell line are known as Finite cell line.
• Cell line which are chemically or virally transformed (transformation)
acquires the ability to proliferate indefinitely, these cell line is called
continuous cell line.
• Contact inhibition and anchorage dependence and low growth rate are the
characteristics of finite cell line.
• Absence of contact inhibition and anchorage dependence and high growth
rate are some attributes for continuous cell line.
• Finite cell line grow as monolayer culture i.e cells cover the bottom of the
culture vessel uniformally.
• Continuous cell line may be grow as monolayer or suspension culture (cells
do not adhere but remain suspended in culture media).
11. DMEM MEDIA COMPOSITION
Note- 10% DMEM media is required
for normal cell culture like Vero or
C6/36 or BHK-21 cell lines.
✓ So to make 10%, we have to add
FBS and streptomycin/penicillin
solution, now we have complete
DMEM MEDIA which is ready to
use for culturing
✓ All other components are already
added in media so no need to add
others.
12. Cell culture incubator
Major equipments required for cell
culture
Bio Safety cabinet
Sub-culturing of cells on confluency
16. Things to be done before starting cells passaging
1) First of all we have to give UV radiation to culture hood for sterilisation.
2) Media which was kept in 4°C should be remain on room temperature
before start.
3) Clean the Hood with 70% ethyl alcohol.
4) Then keep all necessary things in culture hood after rinsing them with
alcohol.
5) Discard should be there in hood to discard the spent media of culture
flask.
6) Check the confluency of cells first under microscope. Cells should be
healthy.
7) Never split the cells which are not confluent. Otherwise it will stress to
the cells. Cells are very fragile and sensitive so handle very gently and
carefully.
8) Always clean your hands with 70% ethyl alcohol.
9) Trypsin/EDTA which is required for cell splitting should be pre-warmed
at 37 °C.
17. Procedure for splitting/passaging of cell line from established
primary cell line (Like Vero cell line which is an adherent cell line)
1) Take the confluent flask and remove all the spent media using serological pipette.
2) Wash the monolayer of cells once with Phosphate buffer saline.
3) Then take 1 mL of Trypsin/EDTA solution and let it cover the monolayer completely.
4) Remove some trypsin solution.
5) Keep the flask at 37°C in BOD incubator for approx. one and half minutes.
6) Tap gently on the side of flask so that cells can dislodge.
7) Add about 2-3 mL of 10% complete DMEM media to flush off the cells that have got dislodged.
8) Centrifuge the tube containing the cells at 1500 rpm for 5 minutes.
9) Cell pellet will form.
10) Now dissolve the pellet in 2 ml fresh DEMEM media.
11) Count the cells by hemocytometer and take the cells accordingly into the new flask to continue the cell
line.
12) Check the flask after 24 hour for its quality and any contamination.
13) Split this flask at alternate day to maintain the cell line for long time (i.e. upto 30 passages).
18. How freezing of cells in done in Liquid nitrogen
• After trypsinisation of Confluent culture flask , forms pellet and
dissolve in DMEM media(as mentioned in previous slide).
• Count the cell to get an idea about cell density.
• Then add 2-3 million cells per cryovial on freezing media.
• Freezing media is basically DMEM media with 10% FBS and 10%
DMSO). DMSO is dimethyl sulphoxide which is a cryoprotectant and
prevent ice crystal formation.
• Keep the cryovial in -80°C and then next day in liquid nitrogen.
• In this way our cell line remains with us for a very long time.
19. Source of contamination in cell culture
• Contamination of cell line means something which is undesirable in the culture system but comes unknowingly.
• Contamination makes our culture media present in cell culture flask fuzzy. Normally it should be clear.
• Two reason may be possible for contamination either
• 1. Chemical contamination
• 2. Biological contamination.
• Chemical contamination includes:-
✓ DMEM media
✓ Phosphate buffer
✓ Trypsin/EDTA solution
✓ Incubator
• Biological contamination includes:-
✓ Bacteria
✓ Fungi
✓ Mycoplasma (generally observed)
✓ Cross contamination with other cell line (i.e When two cell lines are splitted back to back without giving any UV-
radiation, then chances are more).
20. References
• Oyeleye et al (2016). Basics of animal cell culture: Foundation for
modern science. Biotechnology and Molecular Biology Reviews. Vol.
11(2), pp. 6-16, May 2016.
• Yao, T and Asayama, Y. (2016). Animal-cell culture media: History,
characteristics, and current issues. Reprod Med Biol. 2017;16:99.