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W ha t is T is sue Cultur e?

Tissue culture is the growth of tissues and/or cells separate from the organism.
This is typically facilitated via use of a liquid, semi-solid, or solid growth media, such as
broth or agar. Tissue culture commonly refers to the culture of animal cells and tissues,
while the more specific term plant tissue culture is used for plants. Tissue culture is when
a piece of plant is put into the tiniest of greenhouses...a test tube....and a miracle takes
place. If the test tube is free of micro-organisms and the media in the test tube has the
correct balance of chemicals and growth hormones, that little piece of plant will
reproduce replicas or clones of the single parent plant. That little piece of plant material
can replicate in incredible numbers so theoretically, one small piece can produce an
infinite number of clones.

W hy is tissue cultur e used?

Tissue culture is used for many reasons but the 3 major reasons DNA Gardens are
concerned with are 1) Large quantities of plant production for the sake of volume, 2) to
speed up the production of new varieties into the market place. Large amounts of plants
can be produced in a very short time compared to conventional propagation methods, 3)
to establish and maintain virus free stock as in the case of the seed potato industry.
Working in a tissue culture lab is like working in a hospital operating room. Sanitation is
imperative. If the plant cultures become infected with fungus or bacteria, the
contaminants soon over take and kill the cultures. Other applications include protoplast
fusion and the induction and selection of mutants and the biosynthesis of plant
metabolites.
A. Animal T issue s

The development of a fertilized egg into a newborn child requires an average of


41 rounds of mitosis. During this period, the cells produced by mitosis enter different
pathways of differentiation; some becoming blood cells, some muscle cells, and so on.

There are more than 100 visibly-distinguishable kinds of differentiated cells in the
vertebrate animal. These are organized into tissues; the tissues into organs. Groups of
organs make up the various systems - digestive, excretory, etc. - of the body.

TYPE S OF ANIMAL T ISSU ES

1. Epithelial – covering
2. Connective – support
3. Muscular – movement
4. Nervous – control

EPITHELIAL TISSUES
General Characteristics

1. Cellularity – cells are closely-packed with only tiny amount of extra cellular material
in between

2. Specialized contacts – adjacent cells are bound together at many points by lateral
contacts (e.g., gap junctions, tight junctions, etc.)

3. Polarity – have an apical surface (free surface exposed to exterior/cavity) and an


attached basal surface
• Apical surface usually has microvillus or cilia
• Basal lamina is noncellular and adhesive (glycoproteins)

4. Supported by connective tissue – just deep to basal lamina is reticular lamina (layer
of extra cellular material containing collagen fibers); these two lamina form the
basement membrane which reinforces epithelial sheet and defines its boundary

5. Avascular but innervated – nourished by substances diffusing from blood vessels in


underlying connective tissue

6. Regeneration – high regenerative capacity as long as they are nourished

• General functions:

1. Covering and lining


2. Glandular or secretive

Types:

-based on arrangement

1. Simple – composed of single cell layer; located where thin barrier is needed for
absorption or filtration
2. Stratified – composed of 2 or more cell layers stacked on each other; located where
protection is important (areas of high abrasion)

- based on shape in apical layer

1. Squamous- flattened and scale like; nucleus is disc-shaped


2. Cuboidal – boxlike; as tall as they are wide; nucleus is spherical
3. Columnar – tall and column-shaped; nucleus is elongated from top to bottom
and located near base
4. Pseudo stratified – vary in height but all rest on basement membrane; only tallest
reach free surface; cell nuclei lie at different levels giving false impression that
layers are present
• Specific types of epithelial tissue:

1. Simple squamous – cells flattened laterally; sparse cytoplasm; thin layered; found
where filtration or exchange by diffusion is important; can have secretory function

Figure 7B: Schematic representation of simple squamous epithelium.

2. Simple cuboidal – single layer of cells as tall as wide; found where secretion and
absorption is important

3. Simple columnar – single layer of tall cells with oval nuclei; may bear cilia to propel
mucus; may contain mucus secreting unicellular glands (goblet cells); found where
secretion (mucus, enzymes, etc.) and absorption is important.
Figure 4: Schematic representation of simple columnar epithelium.

4. Pseudo stratified – single layer of cells of differing heights; may contain goblet cells
and Bear cilia; found where secretion and absorption is important.

Figure 9A: Schematic representation of pseudostratified epithelium.

5. stratified squamous – several layers thick; free surface cells are squamous while
deeper layers are cuboidal or columnar; can be keratinized (epidermis) or no keratinized
(linings); function in protection from wear and tear underlying tissues.

Figure 8A: Schematic representation of stratified squamous epithelium.


6. Glandular – 1+ cell that make and secrete a product; classified on basis of site of
product release and # of cells (unicellular or multicellular)
• Endocrine – ductless glands that produce hormones; products secreted by
exocytosis directly into extra cellular space; diverse
• Exocrine – secrete products onto body surfaces or into body cavities; products
secreted by exocytosis or via epithelial-lined duct that transports secretion to
surface

CONNECTIVE TISSUE

• General Characteristics:

1. Common origin – all arise from embryonic tissue (mesenchyme)


2. Degree of vascularity – variable; can be avascular, poorly vascularized or richly
vascularized have 3 main elements – ground substance, fibers, and cells

• Ground substance = interstitial fluid, cell adhesion proteins, proteoglycans;


holds large amounts of fluid and functions as molecular sieve.
• Fibers = collagen, elastic, reticular fibers
• Cells = fibroblasts (connective tissue proper), chondroblasts (cartilage),
osetoblasts (bone), accessory cells (WBCs, mast cells, and macrophages), blood
cells
4. Extra cellular matrix – nonliving; separates living cells from non-living; makes tissue
more capable of bearing weight and withstanding tension (= ground substance and
fibers)
• General functions:
1. Binding and support
2. Protection
3. Insulation
4. Transportation of substances

• Specific types of connective tissue:

1. Areolar tissue (loose) – functions include supporting and binding other tissues, holding
body fluids, defending against infection, and storing nutrients and water (holds as much
water as in bloodstream)
• Ground substance is semi fluid or gelatinous
• Fibers = all 3 types but loosely arranged
• Cells = fibroblasts, macrophages, fat cells, mast cells, WBCs

2. Adipose tissue – functions include providing reserve food fuel, insulation, support and
protection of organs; richly vascularized b/c of high metabolic activity
• Ground substance is semi fluid or gelatinous, but scanty so cells are packed
closely
• Fibers = all 3 types but sparsely arranged
• Cells = adiposities, fat cells predominate and are packed closely together giving
a chicken-wire appearance to tissue; oil droplets occupy most of the volume;

3. Reticular – forms internal framework that can support many free blood cells in lymph
nodes, spleen, and bone marrow
• Ground substance is loose
• Fibers = only reticular fibers
• Cells = fibroblasts (reticular cells) are scattered on fiber network

4. Dense fibrous – fibers are primary component of tissue

A. dense regular connective tissue – forms tendons (attach muscle to bone),


aponeuroses (attach muscle to other muscle or bone), ligaments (bind bones)
• Ground substance is scant
• Fibers = collagen fibers mainly, some elastic fibers; all run in same direction
and in direction of pull
• Cells = fibroblasts in rows along fibers

B. dense irregular connective tissue – found in areas where tension is exerted in many
directions at once, such as skin (dermis), joint capsules, fibrous coverings of testes,
kidneys, bones, cartilages, muscles, and nerves
• Ground substance is scant
• Fibers = thick bundles of collagen fibers, irregularly arranged
• Cells = fibroblasts in rows along fibers

5. Cartilage – avascular and aneural


• Ground substance has large amounts of glycosaminoglycans, making it more
stiff and lots of tissue fluid (up to 80% water)
• Fibers = collagen and elastic fibers
• Cells = chondrocytes and chondroblasts (immature cells)

A. hyaline cartilage (gristle) – lots of collagen fibers and few chondrocytes; provides
firm support and some pliability

B. fibro cartilage – rows of chondrocytes alternate with rows of thick collagen fibers;
compressible to resist tension
C. elastic cartilage – more elastic fibers to that it has both strength and stretchability.

6. Bone – functions in support and protection, provides levers for muscles to act on,
stores calcium and other minerals and fats; marrow is site of RBC formation
• Matrix is similar to cartilage but more rigid b/c of presence of inorganic
calcium
salts; salts are deposited on and between fibers
• Cells = osteoblasts that produce the organic matrix

7. Blood – functions as transport vehicle for cardiovascular system (carries nutrients,


wastes, gases, etc. throughout body)
• Matrix is blood plasma (nonliving fluid) + soluble protein molecules that can
form fibers (visible only during clotting)
• Cells = RBCs

MUSCLE TISSUE
• General characteristics:
- Highly cellular
- Well-vascularized
- possess myofilaments (composed of actin and myosin molecules) that bring about
movement

• General functions:

According to types:

1. Skeletal (voluntary muscle) – packaged by connective tissue into sheet-like organs;


have long, cylindrical, multinucleate cells; have striations reflective of alignment of
Myofilaments

2. Cardiac (involuntary muscle) – found only in wall of heart; cells are uninucleate and
Branching and fit together at junctions called intercalated discs; have striations

3. Smooth (involuntary muscle) – spindle-shaped cells with centrally located nucleus; no


obvious striations; found in walls of hollow organs; functions to squeeze substances
through organs

NERVOUS TISSUE

• General characteristics
- makes up brain, spinal cord, and nerves

• General functions:

1. Transmit electrical signals from sensory receptors to effectors (muscles and glands)
• Cell types:
• Neurons – highly specialized nerve cells that generate and conduct nerve
impulses; are branching cells
• support cells (neuroglial cells or glial cells) – non-conducting cells that support,
insulate and protect neurons.

Wound Healing - repair of tissues


• Regeneration - destroyed tissues replaced with tissues of same kind
• Fibrosis - destroyed tissues replaced by fibrous connective tissue

BODY MEMBRANES – comprised of both epithelial and connective tissue

A. Cutaneous (skin) – consists of keratinized stratified squamous epithelium attached to


a thick layer of dense irregular connective tissue (dermis); is a dry membrane
B. Mucous – consists of either stratified squamous or simple columnar epithelium
attached to a layer of loose connective tissue called lamina propria, which may in turn sit
on a layer of smooth muscle; is a wet membrane (secretes mucus) that lines all body
cavities that open to the exterior
C. Serous – consists of simple squamous epithelium resting on areolar connective tissue;
is a wet membrane (secretes serous fluid) that lines all body cavities (peritoneum, pleural,
Pericardial) that are closed to the exterior; has 2 layers
• Parietal – pertains to the walls of the cavity (outermost layer)
• Visceral – pertains to outer wall covering of internal organ or inner part of
structure.

B . P lant Tissue s

Plants are composed of three major organ groups: roots, stems and leaves. As we
know from other areas of biology, these organs are comprised of tissues working together
for a common goal (function). In turn, tissues are made of a number of cells which are
made of elements and atoms on the most fundamental level. In this section, we will look
at the various types of plant tissue and their place and purpose within a plant. It is
important to realize that there may be slight variations and modifications to the basic
tissue types in special plants. Plant tissues are characterized and classified according to
their structure and function. The organs that they form will be organized into patterns
within a plant which will aid in further classifying the plant. A good example of this is the
three basic tissue patterns found in roots and stems which serve to delineate between
woody dicot, herbaceous dicot and monocot plants. We will look at these classifications
later on in the tutorial.
Types o f Plant Tissues

Examples of tissue in other multicellular organisms are vascular tissue in plants, such
as xylem and phloem. Plant tissues are categorized broadly into three tissue systems: the
epidermis, the ground tissue, and the vascular tissue. Together they are often referred to
as biomass.

• Epidermis - Cells forming the outer surface of the leaves and of the young plant
body.
• Vascular tissue - The primary components of vascular tissue are the xylem and
phloem. These transport fluid and nutrients internally.
• Ground tissue - Ground tissue is less differentiated than other tissues. Ground
tissue manufactures nutrients by photosynthesis and stores reserve nutrients.

Plant tissues can also be divided differently into two types:

1. MERISTEMATIC TISSUES
2. PERMANENT TISSUES

Meristimatic tissues

The growth of plant occurs only in certain specific regions. At these regions, the
meristimatic tissues are present. New cells produced by meristem are initially those of
meristem itself, but as they grow and mature, their characteristics slowly change and they
become differentiated as components of other tissues. Depending on the region of
occurrence of meristimatic tissues they are classified as:

a) Apical Meristem - It is present at the growing tips of stems and roots and
increases the length of the stem and root. The girth of the stem or root increases
due to lateral meristem (cambium).This meristem is responsible for the linear
growth of an organ.
b) Lateral Meristem - This meristem consist of cells which mainly divide in one
plane and cause the organ to increase in diameter and girth. Lateral Meristem
usually occurs beneath the bark of the tree in the form of Cork Cambium and in
vascular bundles of dicots in the form of vascular cambium. The activity of this
cambium results in the formation of secondary growth.
c) Intercalary Meristem - This meristem is located in between permanent tissues.
It is usually present at the base of node, inter node and on leaf base. They are
responsible for growth in length of the plant.
Characteristics of meristematic tissues

The cells of meristematic tissues are similar in structure and have thin and elastic primary
cell wall made up of cellulose. They are compactly arranged without inter molecular
spaces between them. Each cell contains a dense cytoplasm and a prominent nucleus.
Dense protoplasm of meristematic cells contains very few vacuoles. Normally the
meristimatic cells are oval, polygonal or rectangular in shape.

Permanent tissue

The meristematic tissues that take up a specific role and lose the ability to divide. This
process of taking up a permanent shape, size and a function is called cellular
differentiation. Cells of meristematic tissue differentiate to form different types of
permanent tissue. There are 2 type of permanent tissues:

Simple permanent tissues

These tissues are called simple because they are composed of similar types of cells which
have common origin and function. They are further classified into:

1. Parenchyma
2. Collenchyma
3. Sclerenchyma

Parenchyma

Parenchyma is Greek word where "parn" means besides and "enchien" means to pour.
Parenchyma is the most specialized primitive tissue. It mainly consist of thin-walled cells
which have intermolecular spaces between them. The cell wall is made up of cellulose.
Each parenchymatous cell is iso-diametric, spherical, or oval in shape. It is widely
distributed in various plant organs like root, stem, leaf, flowers and fruits. They mainly
occur in cortex epidermis, pith and mesophyll of leaves.
A line drawing of a parenchyma

Collenchyma

Collenchyma is Greek word where "Collen" means gum and "enchyma" means infusion.
It is a living tissue of primary body like Parenchyma. Cells are thin-walled but possess
thickening of cellulose and pectin substances at the corners where number of cells join
together. This tissue gives a tensile strength to the plant and the cells are compactly
arranged and do not have intermolecular spaces. It occurs chiefly in hypodermis of stems
and leaves. It is absent in monocots and in roots. Collenchymatous tissue acts as a
supporting tissue in stems of young plants. It provides mechanical support, elasticity, and
tensile strength to the plant body. It helps in manufacturing sugar and storing it as starch.
It is present in margin of leaves and resist tearing effect of the wind.

Sclerenchyma
Sclerenchyma is Greek word where "Sclrenes" means hard and "enchyma" means
infusion. This tissue consists of thick-walled, dead cells. These cells have hard and
extremely thick secondary walls due to uniform distribution of lignin. Lignin deposition
is so thick that the cell walls become strong, rigid and impermeable to water.
Sclerenchymatous cells are closely packed without intra-cellular spaces between them.
Thus, they appear as hexagonal net in transverse section. The cells are cemented with the
help of lamella. The middle lamella is a wall that lies between adjacent cells.
Sclerenchymatous cells mainly occur in hypodermis, pericycle, secondary xylem and
phloem. They also occur in endocorp of almond and coconut. It is made of pectin, lignin,
protein. The cells of sclerenchyamtous cells can be classified as:

1. Fibres- Fibres are long, elongated sclerenchyamtous cells with pointed ends.
2. Sclerides- Sclerenchymatous cells which are short and possess extremely thick,
lamellated, lignified walls with long singular piths.
3. They are called sclerides. The main function of Sclerenchymatous tissues is to
give support to the plant.

A line drawing of a sclerenchyma cell.


Complex permanent tissue

A complex permanent tissue may be classified as a group of more than one type of tissue
having a common origin and working together as a unit to perform a function. These
tissues are concerned with transportation of water, mineral, nutrients and organic
substances. The important complex tissues in vascular plants are xylem, phloem.

Xylem

Xylem is a chief, conducting tissue of vascular plants. It is responsible for conduction of


water and inorganic solutes.

1. Tracheids- Trachids are elongated, tube-like dead cells with elongated end-walls.
End walls remain intact and possess piths. In transverse section, They usually
occur as polygonal cells and lignified walls.
2. Vessels - Vessels are placed one upon another. Their end walls are perforated.
They form long tubes or channels for conduction of water and minerals.
3. Xylem Parenchyma - Xylem Parenchymatous cells are living cells present in
xylem. They help in lateral conduction of organic solutes and storage reserves.
4. Xylem Fibres - Xylem Fibres are lignified fibres present in xylem which provide
mechanical strength to the plant body.

Xylem is a major conducting tissue of vascular plants. It serves in upward movement of


water and minerals from root to different parts of plant.
Phloem

Phloem is a chief conducting tissue of vascular plants. It is regarded as a living tissue


responsible for translocation of organic solutes.

1. Sieve tube - Sieve tubes are long tubular structures composed of elongated sieve
tube elements placed one above other forming a continuous tube.
2. Companion cell - Companion cells are living cells always associated with sieve
tubes. Sieve tube elements and companion cells arrive from the same, initial cell
and therefore forms a single functional unit. Each companion cell shows presence
of fine piths with all the living components of the cell.
3. Phloem Parenchyma - These cells are living parenchymatous cells that are
present in phloem. These cells help in storage of food.
4. Phloem Fibres - Phloem fibres are formed by dead, sclerenchymatous fibres.

The main function of phloem is translocation of organic solutes from the leaves to the
storage organ and later from the storage organ to the growing part. Sieve tube allow free
diffusion of soluble, organic substances across sieve plates due to the presence of large
number of sieve pores.