The document discusses stem cell culture, detailing its history, characteristics, and differentiation, as well as the various types of stem cells including embryonic and adult stem cells. It outlines the process of culturing embryonic stem cells in a laboratory setting, their applications in medical treatments, and the technical challenges associated with stem cell research. The conclusion emphasizes the significance of stem cells in medical advancements and their role in tissue repair and genetic studies.

1. STEM CELL CULTURE
By
KAUSHAL KUMAR SAHU
Assistant Professor (Ad Hoc)
Department of Biotechnology
Govt. Digvijay Autonomous P. G. College
Raj-Nandgaon ( C. G. )

2. Content
 Introduction
 History
 Stem cell characteristic
 Stem Cell Differentiation
 Kinds of stem cell
 Embryonic stem cell
 Stem cell application
 Technical challenges
 Conclusions
 References

3. Introduction
 Cell culture is the process by which prokaryotic,
eukaryotic or plant cells are grown under controlled
conditions. But in practice it refers to the culturing of
cells derived from animal cells.
 Roux in 1885 for the first time maintained embryonic
chick cells in a cell culture.
 Cell culture was first successfully undertaken by Ross
Harrison in 1907.

4. 5/15/2020
1998 - Researchers first extract stem cells from human embryos
1999 - First Successful human transplant of insulin-making cells
from cadavers
2001 - President Bush restricts federal funding for embryonic
stem-cell research
2002 - Juvenile Diabetes Research Foundation International
creates $20 million fund-raising effort to support stem-cell
research
2004 - Harvard researchers grow stem cells from embryos using
private funding
2004 - Ballot measure for $3 Billion bond for stem cells
Stem Cell History

5. Stem Cell – Definition
 A cell that has the ability to continuously divide and
differentiate (develop) into various other kind(s) of
cells/tissues
ES cells from mouse embryos have been cultured since
the 1980s by various groups of researchers working
independently

6. 5/15/2020
Stem Cell Characteristics

7. 5/15/2020 Dr. Hariom Yadav
Stem Cell Differentiation

8. 5/15/2020
Kinds of Stem Cells
Embryonic stem cells come from a five
to six-day-old embryo. They have the
ability to form virtually any type of cell
found in the human body.
Embryonic germ cells are derived from
the part of a human embryo or foetus that
will ultimately produce eggs or sperm
(gametes).
Adult stem cells are undifferentiated cells
found among specialised or differentiated
cells in a tissue or organ after birth.

9. Embryonic stem cell
 Embryonic stem cells (ES cells)
are pluripotent stem cells
derived from the inner cell mass
of a blastocyst, an early-stage
embryo.
 Human embryos reach the
blastocyst stage 4–5 days post
fertilization, at which time they
consist of 50–150 cells.

10. How are embryonic stem cells grown in the
laboratory?
 Growing cells in the laboratory is known as cell culture.
 Human embryonic stem cells are isolated by transferring the inner cell
mass into a plastic laboratory culture dish that contains a nutrient broth
known as culture medium.
 The inner surface of the culture dish is typically coated with mouse
embryonic skin cells that have been treated so they will not divide. This
coating layer of cells is called a feeder layer.

11. Continue….
The reason for having the mouse cells in the bottom of the culture dish is to
give the inner cell mass cells a sticky surface to which they can attach. In
addition, these feeder cells release nutrients into the culture medium.
Over the course of several days, the ESC proliferate and begin to crowd the
culture dish.
Cells are removed gently and plated into several fresh culture dishes. The
process of replating the cells is repeated many times and for manymonths,
and is called subculturing. Each cycle of subculturing the cells is referred to
as a passage.

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Stages of Embryogenesis
Day 1
Fertilized egg
Day 2
2-cell embryo Day 3-4
Multi-cell embryo
Day 5-6
BlastocystDay 11-14
Tissue Differentiation

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Derivation and Use of
Embryonic Stem Cell Lines
Isolate inner cell mass
(destroys embryo)
Heart muscleKidney
Day 5-6
Blastocyst
Inner cells
(forms fetus)
Outer cells
(forms placenta)
Heart
repaired
Culture cells

14. 5/15/2020 Dr. Hariom Yadav

15. 5/15/2020 Dr. Hariom Yadav

16. Stem Cell Applications
Tissue repair
- nerve, heart, muscle, organ, skin
Cancers
Autoimmune diseases
- diabetes, rheumatoid arthritis, MS
Production of transgenic animals
In surgical repair and tissue grafting
Drug discovery and development
Study of biological processes

17. Tissue Repair
• Regenerate spinal cord, heart tissue or any other major in
the body.

18. Heart Disease
• Adult bone marrow stem cells injected into the hearts
are believed to improve cardiac function in victims of
heart failure or heart attack

20. Leukemia and Cancer
• Studies show leukemia patients treated with stem
cells emerge free of disease.
Injections of stem cells have also reduces
pancreatic cancers in some patients.
Proliferation of white cells

21. Rheumatoid Arthritis
• Adult Stem Cells may be helpful in jumps tarting repair
of eroded cartilage.

22. Type I Diabetes
• Pancreatic cells do not produce insulin
Embryonic Stems Cells might be trained to become pancreatic
islets cells needed to secrete insulin.

23. Technical Challenges
 Recently, abnormalities in chromosome number and
structure were found in three human ESC lines.
 Stem cell development or proliferation must be
controlled once placed into patients.
 Possibility of rejection of stem cell transplants as
foreign tissues is very high.
 Contamination by viruses, bacteria, fungi, and
Mycoplasma possible.
 The use of mouse “feeder” cells to grow ESC could
result in problems due to xenotransplantation.

24. Why is Stem Cell Research So Important
to All of Us?
Stem cells can replace diseased or damaged cells
Stem cells allow us to study development and genetics
Stem cells can be used to test different substances (drugs and
chemicals)

25. Conclusion
 Stem cells are biological cells found in all
multicellular organisms, that can divide (through
mitosis) and differentiate into diverse specialized.
 In adult organisms, stem cells and progenitor cells
act as a repair system for the body, replenishing
adult tissues.

26. References
 CULTURE OF ANIMAL CELLS 5TH EDITION : BY R. IAN
FRESHNEY.
 BIOTECHNOLOGY : U. STAYANARAYAN REPRINT 2009