8 Jun 2011

ANIMAL CELL CULTURE


Salient Features of Animal cell culture

A) Animal cells can grow in unadorned schooner or plastic containers in nutritive media but they grow single to restricted generations.

B) Animal cells exhibit friend inhibition.Inside culture the cancer cells apparently clash from the habitual cells. Due to abandoned growth and more rounded affect, they baggy friend inhibition and heap ended all other.

C) There is a difference in the in vitro and in vivo growth pattern of cells.
For model
(i) here is an absence of cell-cell interaction and cell matrix interaction,
(ii) here is a lack of three-dimensional architectural advent, and
(iii) altered hormonal and nutritional background. They way of adherence to schooner or plastic container in which they grow, cell proliferation and affect of cell results in alterations.

D) The maintenance of growth of cells under laboratory conditions in apposite culture standard is renowned as PRIMARY CELL CULTURE.

E) Cells are dissociated form tissues by mechanical earnings and by enzymatic incorporation using proteolytic enzymes.

F) Cells can grow as adherent cells (anchorage dependent) or as suspension cultures (anchorage independent).

G) The primary culture is subcultured in fresh media to set up SECONDARY CULTURES.

H) The various types of cell shape are categorized into two types as Finite cell line and Continuous cell line. Finite cell shape are persons cell shape which be inflicted with a restricted life span and grow through a restricted digit of cell generations. The cells normally divide 20 to 100 era (i.E. Is 20-100 population doublings) previous to extinction. Cell shape transformed under in vitro conditions produce increase to unremitting cell shape. The unremitting cell shape are transformed, immortal and tumorigenic.

I) The corporal background includes the optimum pH, warmth, osmolality and gaseous background, at the bottom of go up and caring the cells from compound, corporal, and mechanical stresses.

J) Nutrient media is the mixture of inorganic salts and other nutrients competent of sustaining cell survival in vitro

K) Serum is essential pro creature cell culture and contains growth factors which promote cell proliferation. It is obtained as exuded liquid from blood undergoing coagulation and filtered using Millipore filters.

L) Cryo maintenance is storing of cells by very low warmth (-1800C to -196 0C) using liquid nitrogen. DMSO is a cryopreservative molecule which prevents destruction to cells.

M) inside order to keep up the aseptic conditions in a cell culture, a LAF covering is used. Based on the nature of cells and organism the tissue culture hoods are grouped into three types: Class I, Class II, and Class III.

N) CO2 incubators are used and designed to mimic the environmental conditions of the living cells.

O) An inverted microscope is used pro visualizing cell cultures in situ

P) For generally creature cell cultures low alacrity centrifuges are looked-for.

Q) Neuronal cells constitute the worried logic. Inside culture the neuronal cells cannot divide and grow.

R) The cells with the intention of form connective tissue (skin) is called fibroblast. The fibroblast can divide and grow in culture to approximately generations with which they go to meet your maker. All habitual creature cells are mortal.

S) Organ culture- The culture of native tissue with the intention of retains generally of the in vivo histological facial appearance is regarded as organ culture.

T) Histotypic culture- The culturing of the cells pro their reaggregation to form a tissue-like organize represents histotypic culture.

U) Organotypic culture- This culture practice involves the recombination of uncommon cell types to form a more defined tissue or an organ.

There are particular stipulations with the intention of are associated with the cell shape.
These are as follows:
(i) Split ratio- The divisor of the dilution ratio of a cell culture by subculture.
(ii) Passage number- It is the digit of era with the intention of the culture has been educated,
(iii) Generation number- It refers to the digit of doublings with the intention of a cell population has undergone.

Inside detail these parameters help us to distinguish the cancer cells in culture from the habitual cells since the cancer cells in culture, exchange affect (more rounded), baggy friend inhibition, heap on all other due to overgrowth and abandoned growth.

REQUIREMENTS FOR ANIMAL CELL CULTURE

Among the essential supplies pro creature cell culture are special incubators to keep up the levels of oxygen, carbon dioxide, warmth, damp as bestow in the animal’s body. The phony media with vitamins, amino acids and fetal calf serum. Following parameters are essential pro thriving creature cell culture:

A) Temperature- inside generally of the mammalian cell cultures, the warmth is maintained by 370C in the incubators as the body warmth of Homo sapiens is 370C.

B) Culture media- The culture media is prepared in such a way with the intention of it provides-

    1) The optimum conditions of factors like pH, osmotic pressure, and that.

    2) It must contain compound constituents which the cells or tissues are incapable of synthesizing. Generally the media is the mixture of inorganic salts and other nutrients competent of sustaining cells in culture such as amino acids, full of fat acids, sugars, ions, trail elements, vitamins, cofactors, and ions. Glucose is added as energy source-it’s concentration unreliable depending on the requirement. Phenol Red is added as a pH indicator of the standard.There are two types of media used pro culture of creature cells and tissues- the natural media and the synthesized media.

    3) Natural Media - The natural media are the natural sources of nutrient sufficient pro growth and proliferation of creature cells and tissues. The Natural Media used to promote cell growth fall in three categories.

        I) Coagulant, such as plasma clots. It is currently commercially unfilled in the form of liquid plasma reserved in silicon ampoules or lyophilized plasma. Plasma can furthermore be prepared in the laboratory taking made known blood from male fowl and count heparin to prevent blood coagulation.

        Ii) Biological fluids such as serum. Serum is lone of the very valuable components of creature cell culture which is the source of various amino acids, hormones, lipids, vitamins, polyamines, and salts containing ions such as calcium, ferrous, ferric, potassium and that. It furthermore contains the growth factors which promotes cell proliferation, cell attachment and hold factors. Serum is obtained from creature adult blood, placental, cord blood, horse blood, calf blood. The other forms of biological fluids used are coconut fill up, amniotic fluid, pleural fluid, insect haemolymph serum, culture dregs, aqueous humour, from eyes and that.

        Iii) Tissue extracts pro model Embryo extracts- Extracts from tissues such as kernel, liver, pique, leukocytes, tumour, bone spirit and that are furthermore used pro culture of creature cells.

        Synthetic media

        Syntheic media are prepared artificially by count several organic and inorganic nutrients, vitamins, salts, serum proteins, carbohydrates, cofactors and that. Different types of phony media can be prepared pro a variety of cells and tissues to be educated. Synthetic media are of two types- Serum containing media (media containing serum) and serum- emancipated media (media with made known serum). Examples of approximately media are: Smallest essential standard (MEM), RPMI 1640 standard, CMRL 1066, F12 and that.

        Advantages of serum in culture standard are:
        I) serum binds and neutralizes toxins,
        (ii) serum contains a complete fit of essential growth factors, hormones, attachment and diffusion factors, binding and convey proteins,
        (iii) it contains the protease inhibitors,
        (iv) it increases the buffering room,
        (v) it provides trail elements.

        Disadvantages of serum in culture standard are:
        (i) it is not chemically defined and therefore it’s arrangement varies a ration,
        (ii) it is now and again source of contamination by viruses, mycoplasma, prions and that,
        (iii) it increases the difficulties and cost of down spill dispensation,
        (iv) it is the generally expensive element of the culture standard.

    4) pH- Most media keep up the pH linking 7 and 7.4. A pH not more than 6.8 inhibits cell growth. The optimum pH is essential to keep up the proper ion balance, optimal functioning of cellular enzymes and binding of hormones and growth factors to cell go up receptors in the cell cultures. The parameter of pH is made using a variety of buffering systems. Most media aid a bicarbonate-CO2 logic as its major element.

    5) Osmolality- A exchange in osmolality can affect cell growth and function. Salt, Glucose and Amino acids in the growth media determine the osmolality of the standard. All money-making media are formulated in such a way with the intention of their final osmolality is around 300 mOsm.

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