4TH QUARTER

STEM CELLS
• Stem cell research - has attracted the attention of the public both because of the potential for
human health and because of the ethical implications.

SOMATIC CELL, GERM CELLS, STEM CELLS, ADULT STEM CELLS, AND EMBRYONIC STEM CELLS
• Cells – are considered either somatic or germ cells.
• Somatic cells - compose the tissues of our bodies, contain two copies of each chromosome, and
are known as diploid (two copies of everything).
- are typically highly differentiated or grown up.
• Germ cells - are either egg or sperm. They contain only one copy of each chromosome and are
known as haploid (one copy of everything).
• Differentiated cells - usually highly specialized cells and are fully developed. They are committed
to their role in life, and their appearance and cellular chemistry are devoted to their particular
function.
• Undifferentiated cells - are more primitive and do not perform high-level functions.
• Stem cells - These undifferentiated cells retain the capability to specialize, to chose an
occupation if you will.
- they have the ability to reproduce indefinitely. They also retain their ability to
differentiate into specialized cells.
- it can become monocytes, lymphocytes, neutrophils, basophils, and erythrocytes.
• Hematopoietic cells - Cells that give rise to blood cells
• Hematopoietic stem cells (HSCs) - stem cells found in bone marrow.
• Adult stem cells - Stem cells existing in tissues
• Embryonic stem cells (ESCs) - stem cells derived from embryos
- come from fertilized eggs
- diploid
- are obtained early enough in the life of the zygote that they retain
the capability to become any of the types of cells needed by the complete organism
• Zygote - When an egg is fertilized, it divides to form a mass of cells
• Totipotent - it can develop into the entire complex organism and hence
• Multipotent - embryonic stem cell cannot to be induced to form an intact organism, at least not
under current technology
• Pluripotent - devoted to become cells of a particular organ. For example, the HSCs can become
any of the cells of the bone marrow, but their potential to become liver or kidney has yet to be
demonstrated

ADULT STEM CELLS

• Adult stem cells - have been derived from adult tissues, from umbilical cords, and even from
fetal tissues. Note that all of these are considered adult stem cells even though their sources are
not necessarily adult tissues.
- currently the only type of stem cells routinely used to treat human diseases.
 • Stem cells - stem cells have the capability to become a wider range of cells than previously
thought.
• HSCs - have been injected into mice that have widespread tissue damage due to exposure to
high levels of radiation
- have been reported to migrate into tissues other than bone marrow and repair damaged
lungs, livers, and kidneys.
• Stem cells in bone marrow transplant - used to treat victims of blood cancers and also of
hemophilia.
• Stem cells - are important in the growth of skin grafts for plastic surgery for severe burn victims.
- may secrete growth promoting factors and contribute to the formation of new
blood vessels. Stem cells from fatty tissue and bone are used to repair cartilage injury in horses.
Stem cells derived from fat tissue are under investigation for breast reconstructive therapy.
- could be induced to develop whole new organs that could then be used to replace
damaged organs.
• Autologous – the result if a portion of the victim’s skin is removed and maintained in culture
where skin stem cells produce additional skin and enlarge the original tissue.

EMBRYONIC STEM CELLS

• Embryonic stem cells - offer a huge advantage over adult stem cells in that they can be grown in
culture.
- are simply cells of an early stage embryo.
- could develop into the entire organism, although this has never been
demonstrated
• Blastocyte - a ball of approximately 150 cells.

DEVELOPING EMBRYONIC STEM CELL LINES

• Embryonic cells - are harvested from donated eggs, usually those that were fertilized as part of
an in vitro fertilization (IVF) process.
- are “immortal,” that is, they will grow and divide indefinitely
• IVF - used to enhance fertility by removing sperm from the male and egg from the female,
allowing fertilization to occur outside of the body, and then implanting some of the resulting
zygotes into a receptive uterus, usually that of the egg donor.
- increasingly being used for reasons other than remediation of problems with fertility, such
as to allow sex selection.
- used to produce a cattle
• First IVF baby – was born in 1978 in England.
• Frozen zygote - can be transported long distances before implantation into a cow much more
conveniently than the calves could be moved after birth by their natural mother
• up to eight eggs are fertilized per session, and only two or so of these will be implanted into the
prospective mother
• 400,000 - frozen embryos in the United States.
• 2.8% - percentage of frozen embryos in US that has been donated to research.
• Cell line - is kind of like the sourdough in your refrigerator.
• Human ESCs - have undergone 300–400 duplications.
• Mouse ESCs - have existed in culture for several decades.
 • Human fibroblasts - types of cell lines that have been maintained in culture for long periods of
time.
• Human embryonic stem cell (HESC) lines - very fragile, so establishing them is quite a feat.
• HESCs ball up into what are called embryonic bodies and proceed to differentiate rather
randomly.
▫ If injected into mice at this stage, they form a type of tumor called teratoma.
• Teratomas - are tumors that sometimes develop in the uterus after the death of a fetus.
• HESC ball - does not differentiate in an orderly fashion to produce an organism is evidence that
somehow the orchestration of cell differentiation has degenerated in the embryonic stem cell
culture.
• Feeder cells - is a bit of problem when you are intent on injecting HESCs into people for
therapeutic purposes

THERAPEUTIC USES OF EMBRYONIC STEM CELLS

• research using embryonic stem cells is very, very new.
• diseased animals - have improved when their hearts are injected with embryonic stem cells
• Mouse embryonic stem cells - have been studied for decades, and methods to force their
differentiation into various types of tissues are well understood.
• Mouse studies - have confirmed that ESCs can alleviate the symptoms of diabetes, Parkinson’s
disease, and spinal cord injury.
• Geron - announced plans to apply for clinical trials for neurological diseases
- owns the patents for 9 of the 22 embryonic cell lines used in federally funded research.
• Stem cells - are postulated to be the source of at least some cancers in vivo.
• HESCs - may need to employ cells that have been induced to partially differentiate.
- could be genetically engineered so that they do not express cell surface antigens, thus
reducing their antigenicity
• ESCs - are antigenically very different from the patient, the patient’s immune system will seek
and destroy them.
- are reputedly less antigenic than their adult counterparts.

SOMATIC NUCLEAR TRANSFER – CLONING

• Somatic cell nuclear transfer (SCNT) - a method of developing a cell that behaves like an
embryonic stem cell but contains the genetic material derived from an adult cell. The process
begins with a fertilized egg and a somatic cell from an adult.
- it carries some caveats. Animals cloned using SCNT
experience a high rate of late-term miscarriages and severe birth defects, indicating that the cell
transformation is not without flaws.
• Nucleus – it is removed from the fertilized egg and is replaced by the nucleus from the somatic
cell.
• Somatic cell - can be theoretically any type of cell, from bone marrow to skin.
• Egg - is induced to develop into a zygote and an embryonic cell line is derived from the
blastocyte, just like with any embryonic stem cell line development.
• Dolly the sheep - the first successful clone derived from adult cells.
 • February 2004 - a group led by veterinarian Woo Suk Hwang and gynecologist Shin Yong Moon
of Seoul National University shocked the scientific world by reporting the first derivation of ES
cells from human nuclear transfer experiments (Science, 12 March 2004, p. 1669).
• Clone - would be developed only if the modififi ed germ cell (egg) were to be implanted in the
uterus and allowed to develop into a baby that is genetically identical to the individual who
donated the somatic cell.
• Embryonic cell line - could be developed using somatic cells from an individual possessing a
diseased organ.
• Embryonic cells - could be induced to form a completely new organ and this organ transplanted
into the patient.
•

CONTROVERSY AND LEGAL CONSTRAINTS

• U.S. Congress - banned the use of federal funding for embryo-destroying research
• Dr. James Thomson - The first embryonic stem cell line in the United States was therefore
established with private funding and was done in 1998 by a group led by him at the University of
Wisconsin.
The research was funded by the Wisconsin Alumni Research Foundation (WARF) and Geron

• Federal funds to support HESC research were established under the George Bush
administration and have only been available since August 9, 2001.
• There are 22 human embryonic stem cell lines that meet this federal criterion.
• Arizona and Pennsylvania have decreed that the creation of a human embryonic stem cell line
is a felony.
• Of the approximately 130 HESCs worldwide, 70 are owned by U.S. companies or universities.
• The 22 approved cell lines contain the genomes of 22 individuals.
• Wisconsin Alumni Research Foundation - has a patent for “a method of culturing human
embryonic stem cells and composition of matter which covers any cells with the characteristics
of stem cells.”
• A model might be the National Institute of Health’s Recombinant DNA Advisory Committee
(RAC).
• RAC - was formed because of the concerns of both the public and the scientific community
when recombinant DNA technology was first developed that this technology might be misused.
- is a panel of up to 21 national experts in various fields that advises the NIH Director and
the NIH Office of Biotechnology Activities (OBA).
- reviews all research proposals involving human gene transfer.

• Stem cells - can serve as the source of other more highly differentiated cells.
• Adult stem cells - have been isolated from several body tissues and exist apparently to provide a
repair mechanism for a specific organ.
- have been used clinically in bone marrow transplants and to aid in forming skin
grafts.
 • Human embryonic stem cells (HESC) - may differentiate into many cells crossing tissue types
- have enormous therapeutic potential as demonstrated in
animals.
• Somatic cell nuclear transfer - is the process by which the nucleus of an embryonic cell is
replaced by the nucleus of a somatic cell. The embryonic cell then becomes a clone of the
somatic cell and forms embryonic stem cells.
- the process used to clone animals and has the potential to
produce embryonic stem cells that are genetically identical to the individual who might benefit
from therapeutic uses of these cells

INDUSTRIAL AND ENVIRONMENTAL APPLICATIONS

• Microbes - can produce most carbon-based structures that we need.
• Bioproduction – it will compete with petrochemical methods as the most economically feasible
way to obtain the organic chemicals that we depend upon.
• Cell – an exquisite set of manufacturing, destruction, and demolition processes— very much like
a manufacturing plant.
• Cellular manufacturing process – the purpose of this process are to extract energy from the
environment and use this energy to sustain cellular existence, to ward off invaders, and to
reproduce.
• Enzymes – are a subset of proteins—proteins that act as catalysts.
- typically accelerate biochemical reactions by 10^10 (Salsh, 2001), a feat that you
could not expect from your average organic chemist.
- include catalysts that can break down other proteins and catalysts that can serve as
switches, moving chemical reactions in different directions.
• Catalyst - molecular chemists that speed up or alter chemical reactions.

USE OF BIOREAGENTS IN INDUSTRY

• Bioreagents – chemicals that are captured from living cells and used as chemical reagents in
production processes, either within the cell or outside of the cell.
- current widespread of bioreagents can be found in the textile and food industry,
and in pharmaceuticals.
• Low-calorie sweetener aspartame – produced on a kiloton scale by Holland Sweetener
Company using a proteolytic enzyme, thermolysin.
• Industrial applications – it includes the production of phenolic resins, acrylamide, and many
modern insecticides.
• Natural enzymes – that are included in detergents come from cellular enzymes.

ENVIRONMENTAL APPLICATIONS – HAZARDOUS WASTE

• Many groundwater “pump-and-treat” processes strip the contaminants from the water and
release them into the air.
• High temperature incineration – breaks down the contaminant but is very expensive and can
produce toxic air emissions.
• Bioremediation – offers another option: the use of microbes at the site or at the hazardous
waste treatment plant or landfill to convert chemical compounds into innocuous or less harmful
 chemical compounds.
- it is even an option with metal contamination.
• Biostimulation – refers to methods to jump-start the microbes that are already in place by
fertilizing with nitrogenous and other nutritious compounds.
• Bioavailability – an index of how accessible a given chemical is to the plants and microbes in an
area.
• Meal-eating bacteria – it occurs naturally in soil.
• Among the most famous of the metal-eating bacteria are anaerobic bacteria that reduce
hexavalent chromium to trivalent chromium.
• Hexavalent chromium – causes cancer in humans where trivalent does not.
• Chromium-contaminated soil – is dug up and treated in a bioreactor so that conditions for
microbial action can be optimized.
• Some contaminants require a consortium of microbes, for example, polyaromatic hydrocarbons
and PCBs. Treatment of these sites requires bioaugmentation, or introduction of new microbes.

ENVIRONMENTAL APPLICATIONS – AIR EMISSIONS

• These technologies have been applied in principle to emissions that range from low
concentration of contaminants to flue gases containing sulfur and nitrogen. Methods include
use of biofilters or of bioscrubbers, where the gas is sprayed with liquids containing the
microbe.
• Biofilters - consist of a support medium, such as sheets of plastic, to which the microbes adhere,
and a method to bathe the medium with aqueous nutrients.

MARINE BIOTECHNOLOGY
• Office of technology assessment – defined marine biotechnology in 1991 as “any techinique
that uses living-marine organisms (or parts of these organisms) to make or modify products, to
improve plants or animals, or to develop microorganisms for specific uses.”
• 100 million – tons of seafood that is harvested each year to feed a growing world population.
• Fish – main protein staple for more than one billion people in Asia alone.
- may play a larger role in feeding this population
• Oceans – an ancient ecosystem where life, in the form of bacteria, originated about 4 billion
years ago.
• Marine organisms – provide us with a valuable gene – pool that is beginning to be tapped.
- have unique metabolic pathways and other adaptive functions.
• 2% - of federal funds devoted to biotechnology have been remarked for marine biotechnology
and aquaculture.

AQUACULTURE
• Aquaculture – the propagation of aquatic animals and plants at high densities in fresh, brackish,
or salt water.
- has been practiced throughout the world for thousands of years.
 • Salt water – also known as marine culture or mariculture.
• Ancient aquaculture – which was primarily freshwater, began in the far east; Chinese
aquaculture dates back probably at least 3000 years.
• Penacus monodon – is cultured in ponds at densities of 100,000 to 300,000 prawns per hectare.
• Aquaculture ponds – have the potential of destroying delicate habitats through pollution by
wastes or by decimating.
• Marine freshwater biotechnologies – are used today to increase the yield and quality of finfish,
crustaceans, algae, and various bivalves such as clams and oysters.
• Modern Japanese mariculture – is more productive than freshwater aquaculture and has
recently accounted for more than 92% of Japan's total aquaculture yield.
• The major food products have been the japanese oyster, crassostresa gigas, and a red algae
called nori (porphyra).
• Nori – provides the largest seaweed harvest.
• United Nations food and agricultural organization (FAO) - predicts that by the end of the
century, products from aquaculture will account for 20-25% of the world’s fisheries production
by weight.
- according to their report in 1991, the
wild marine and freshwater and marine aquaculture make up 7% and 5%, respectively, of the
total fish catch.
• Pond humus – can be used to fertilize fields, and livestock waste can serve as pond fertilizer to
stimulate the growth of plankton, which is food for the fish.
• Crop by-products – can be used as fish feed.
• Anaerobic digesters – sometimes used to produce methane gas as an energy source from
aquacultural wastes.

MARINE ANIMAL HEALTH

• Biotechnology – provides opportunities improvement of the health of cultivated aquatic
organism.
• Marine animals – are susceptible to protozoan, bacterial, viral, and fungal diseases.
• Marine pollution – increasing dramatically and is often promotes growth of pathogens (such as
the fungus saprolegnia and the common marine bacterial pathogen vibrio that infect marine
organisms).
• 90% - percentage of marine catch that comes from coastal waters, which are the areas most
susceptible to pollution.
• High densities of animals in aquaculture also in crease the chance of disease. disease
contributes to a significant loss of production each year, with devastating economic
consequences.
• Shrimp aquaculture – it provides an important source of revenue to many Asian and Latin
American countries, primarily through export.
• Taiwan’s production of shrimp has dropped from 114,000 metric tons in 1987 to 50,000 in 1988
and 30,000 in 1991 (of a world total, in 1991, of 690,100 metric tons).
• Methods of bacterial control include the use of anti-bacterial agents such as disinfectants and
antibiotics (tetracycline, chloramphenicol, penicillin, etc.) in ponds or culturing pens.
 • Antibiotic resistance – it could be transferred from bacteria in culturing facilities grow human
bacterial pathogens that cause serious diseases such as typhoid, dysentery, and cholera,
normally treated with antibiotics.
• Antibiotic residues – it could remain in the fish, crustacean, or bivalve consumed by humans.
• Salmon – sometimes become infected with viruses in aquaculture facilities.

TWO MOST DEADLY VIRAL DISEASE

1. Infectious hematopoietic necrosis (IHN) - was first found in 1953, when a tremendous number
of salmon died in Washington state; it has since spread world-wide.
2. Infectious pancreatic necrosis (IPN)
• Protozoans – such as flagellates and ciliates may not kill the fish but can cause extensive
damage by feeding on parts of the fish so that it can no longer be used for food.
• Viral diseases – it poses a particularly difficult problem for fish aquaculture facilties.
• Infected animals – can often become carriers and must be destroyed.
• Efficacious fish vaccines – it would greatly benefit the aquaculture industry, but very few are
available.
• Vaccine (or ung recombinant vaccine against IHN) – is based on a cloned subunit of the viral
protein coat that is produced in bacteria.
- it is being tested in large-scale field trials.
• Marine metabolites and other constituents also may prove to be in-valuable in the fight against
disease.
• Shellfish extract – it has been shown to increase immunity against infection in blue crabs and
prawns as well as to protect eels from Aeromonas infection.
• Biotechnology - can also be used against the spread of disease in aquaculture through the
development of diagnostics.

ALGAL PRODUCTS
• Algae - are a diverse group of photosynthetic eukaryotes that are used throughout the world for
various products, including food.
- are harvested in the wild as well as produced in culture.
- most of the production is from Japan, China, and Korea, although the United States
(especially California, where brown algae, or kelp is harvested).

TWO TYPES OF ALGAE

• Microalgae
• Macroalgae

3 MAJOR GROUPS OF EUKARYOTIC MACROALGAE

1. Green (Chlorophyta)
2. Red (Rhodophyta)
3. Brown (Phaeophyta)
 • Wild seaweeds - were collected for food and medicines as early as 900 BC and are still being
collected in some parts of the world.
• Macrocystis - sometimes called giant kelp because they can grow to 30 m.
• Kelp - was used as fertilizer and as a source of potash and acetone for the production of
explosives.
- has been used for many years as a food supplement and is an important source of
potassium, iodine, and other essential minerals; carbohydrates; and vitamins.
• Kelco company of San Diego - (founded in 1929) was the world’s first producer of algin
products; the first product was kelp meal for livestock feed.
• 70 – approximately number of algin products that are manufactured for many applications.
• Alginates - are the main structural components of the cell wall and intercellular matrix of brown
seaweeds.
- They are used as food thickeners and stabilizers (they retain moisture and assure
smooth texture and uniform thawing of frozen foods).
• Algin – it is added to desserts, dairy products (just check your favorite ice cream), canned foods,
salad dressing, cake mixes, and even beer for foam stabilization.
• Their industrial applications include paper coatings and textile printing, and they are used in
pharmaceuticals (for example, antacids, pill coatings, and capsules) and cosmetics.
• 2$ to 200$ and 250$ to 40000$ - cost of agar and agarose per kilogram, depending on purity.
• Algal cell culture – are in progress to increase the amounts synthesized of such products as
agar.
• Macroalgae, or seaweeds – can be cultured to in vitro by producing protoplast and callus tissue
from which these algae can be regenerated.

• Cell and tissue culture – allows new traits to be selected or genes for specific characteristics to
be transferred.
• Protoplast fusion – allows desirable characteristics from two different organisms to be mixed.
• In vitro propagation and selection – may one day lead to the development of disease
resistance, faster growth, tolerance to variations in light temperature and nutrients and
increased production of metabolites and nutrients.
• Microalgae - comprise a diverse group of both eukaryotic algae (for example green algae) and
proteolytic photosynthesis bacteria (for example cyanobacteria, sometimes called blue-green
bacteria).
- also a source of pigments such as phycoerythrin, phycocyanin, B-carotene, and
zeaxanthin.

5 TYPES OF MICROALGAE THAT HAS BEEN EXPLOITED

• Dunaliella
• Scenedesmus
• Spirulina (a cyanobacterium)
• Porphyridium
• Chlorella

The last three in particular are sources of protein and vitamins.

• Mass culturing - enables large quantities of microalgae to be grown and harvested in outdoor
ponds.
• Arachidonic acid - an essential dietary fatty acid and precursor to prostaglandins and other
important compounds.
• Red unicellular algae porphyridium - has one of the highest concentrations of arachidonic acid-
36% of the total fatty acids. This organism also can be mass cultivated in open ponds.
• Phycobiliproteins - are pigments associated with the photosynthetic apparatus of red algae,
cyanobacteria, and cryptomonads.
• Poryphryidium - an excellent source of phycobiliproteins, because compounds can easily be
extracted, and cells are readily cultured.
• Porphyridium phycobiliproteins - (for example, phycoerythrin in red algae) are a potential
source of phycofluid used to label or tag biologically active molecules such as immunoglobulin,
protein A, and biotin.
• Cyanobacterium - a desirable food source because it readily harvested, its cell wall constituents
are easily digested, and it is approximately 70% protein by dry weight.
- marketed as dried flakes, and is found primarily in health food stores, in fish
food, and in Japanese cuisine.
• The largest culturing facility is near Mexico City, at Lake Texcoco, an ancient lake, now dry, with
an Aztec site.
• Spirulina - propagated in ponds at this lake by pumping underground water into a spiral
evaporation system created by dikes.
• There are culturing facilities in Thailand, Israel, Japan, Taiwan, and the United States; total
worldwide production is approximately 850 tons a year.
• Modern algal biotechnology – involves genetic manipulation, chiefly through mutation and
selection, to produce algae that grow faster in culture, are more disease resistant, synthesize
more of a particular metabolite, and even produce new, unique products.
• Recombinant DNA technology – has not been used extensively, because gene transfer methods
must be established for each type of organism, and specific genes and promoters must be
isolated that would allow the expression of a desired trait or product.
• Marine polysaccharides – have antiviral, anti-ulcer, antitumor, anticoagulant, and cholesterol-
lowering activities.
• Algae – are an efficient, renewable, environmentally friendly source of chemicals, pigments, and
energy.
• Porphyra - generally found in cool, marine waters.
- . It is an important component of shallow marine waters into the intertidal zone.
- It is also a type of 'seaweed' that is commonly eaten by humans and is called 'nori'.
• Nori - a kind of red algae that turns dark green when dried, has been eaten in Japan for more
than two thousand years.
• Red algae porphyra – or nori, has been cultured as a food source in Japan since 1570.
- The culturing process relied on the natural dispersal of spores (actually,
propagules called conchospores) from wild populations in the ocean.
• Brown Algae Undaria (Wakame) and Laminaria (Kombu) - are grown off the coasts of Japan
and China.
 - have a variety of commercial
applications; they are used in noodles, soups, and salads, or with meats.
• Alginic acids (alginates) from brown algae and phycocolloid polysaccharides (agars,
carrageenans) from red algae – were commercially produced early in the twentieth century.
• seventeenth-century France – soda ash was obtained from wild kelp (brown algae).
• mid-nineteenth century – iodine was extracted
• Agar production – originated in China and Japan, probably in the seventeenth century.
• Carrageenans – were produced in the 1830s, first in Ireland and then in the United States.
• alginic acids and phycocolloid polysaccharides - are used in food, industrial products, fertilizer,
and energy production.
• Alginates - are in large demand by several industries: of the more than 35,000 tons produced,
the textile industry uses 50%; foods use 30%; pharmaceuticals use 5%; and paper uses 6%.
• Carrageenan - is used extensively as an extender in foods such as evaporated milk and ice
cream, in toothpaste, and in a variety of cosmetics.
• Agars - are used primarily in foods but also in pharmaceuticals (for example, as a component in
capsules holding medication) and in scientific laboratories for making gels (for example, for gel
electrophoresis) and solidified culture media.

ACRONYMS:
• HSCs – hematopoietic stem cells
• ESCs – embryonic stem cells
• IVF – in vitro fertilization
• RAC – recombinant DNA advisory committee
• HESCs – human embryonic stem cells
• OBA – office of biotechnology activities
• SCNT – somatic cell nuclear transfer
• RDNA – recombinant DNA
• HGH – human growth hormone
• BCG - Bacillus Calmette-Guerin
• CHO – Chinese hamster ovary
• BHK – baby hamster kidney
• USDA – US department of agriculture
• PCBs – polychlorinated biphenyls

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