Draw and label an in vitro cellular aging curve-Phase

Solution Preview

---

Please see the attached file.

G. Draw and label an in vitro cellular aging curve-Phase I, II, and III.

E.Coli IN -VITRO AGING CURVE

H. What are the typical conditions for growing human cells outside the body? Compare use of serum vs. defined medium.

Typical conditions:

 Proper human cell medium are required it may be the basal medium or growth medium

 For different cells different medium are there, so we should choose the appropriate medium to culture

 Content of the medium that is components are very important
for example: Placental Epithelial Basal Medium (500 ml) This basal medium is composed of essential and nonessential amino acids, vitamins, inorganic salts, organic compounds and trace elements. It is used for culturing amniotic epithelial cells when supplemented with Amniotic Epithelial Cell Growth Supplements.

 pH, temperature, incubation period, nutrient level should be proper

Compare use of serum vs. defined medium:

Serum:

The clear, thin and sticky fluid portion of the blood that remains after coagulation is called serum. Serum contains no blood cells, platelets or fibrinogen.

 Animal serum is used as the source of growth factors for microorganisms
 It function as a growth medium for microorganisms
Chemically defined [synthetic] medium:
A defined medium will have known quantities of all ingredients, for microorganisms they consist of providing trace elements, any vitamins required by the microbe and especially a defined carbon source and nitrogen source (for example glucose or glycerol are often used as carbon sources, ammonium salts or nitrates as inorganic nitrogen sources), minimal media are defined media.
a. A chemically defined medium is one prepared from purified ingredients and therefore whose exact composition is known.
b. Not trivial for fastideous:
i. To use a chemically defined medium you must know exactly what your microorganism's growth requirements are.
ii. If this is not known it must be discovered through trial and error and therefore is not a trivial process for fastideous microorganisms.
Serum Defined medium
The clear, thin and sticky fluid portion of the blood that remains after coagulation and it is not a defined one. A defined medium and it has known quantities of all ingredients, so for this we must know microorganism's growth requirements
Based on the serum ingredient, microorganisms will grow Particular ingredients are required to grow

I. Define a population doubling-in vitro. How many can you expect from a human, chicken, and mouse?

Population doubling-in vitro:

Population Doubling in vitro - historically recorded doubling of a given population in in-vitro condition

Population doubling time:

The interval, calculated during the logarithmic phase of growth in which, for example, 1.0 X 106 cells increase to 2.0 X 106 cells.

Population doubling level:

The total number of population doubling of a cell line or strain since its initiation in vitro. A formula to use for the calculation of "population doublings" in a singe passage is:

Number of population doublings = Log10 (N/N0) X 3.33

Where:

N=number of cells in the growth vessel at the end of a period of growth. N0=number of cells plated in the growth vessel.

It is best to use the number of viable cells or number of attached cells for this determination. Population doubling level is synonymous with "cumulative population doublings."

Human:
"It may be safely asserted, therefore, that population, when unchecked, increases in a geometrical progression of such nature as to double itself every twenty-five years. This statement, of course, refers to the general result, and not to each intermediate ...