Various types of cell cultures

various type of cell culture

1. VARIOUS TYPES OF CELL CULTURES 1

2. TYPES OF CELL CULTURES: Cell culture is classified i nto three: Primary cell culture Adherent cell culture Suspension cell culture Secondary cell culture Cell line Finite cell line Continuous cell line 2

3. Primary culture 3 It is the stage of culture after isolation of cells but before the first subculture after which it becomes cell line Four stages Acquisition of the sample Isolation of tissue Dissection/ disaggregation culture

4. 4 Fat and necrotic tissues are best removed during dissection. Tissue should be chopped finely to cause minimum damage. Enzymes used for disaggregation should be removed subsequently by gentle centrifugation.

5. PRIMARY CELL CULTURE : Separate cell directly from the parent tissue ( kidney, liver, heart etc.) Separated by enzymatic or mechanical method. Maintaining the growth of the cell in a culture medium using glass or plastic container. Culture medium Liquid or gel designed to maintain the growth of the cell. It varies from different type of cell. 5

6. Primary cell culture is further classified based on the types of the cells. Adherent cell : Cells shown to require attachment for growth are said to be anchorage dependent cells. The adherent cells are usually derived from tissues of organs such as kidney where they are immobile and embedded in connective tissue. Those cell which attach to the surface of the culture flask. Forms a monolayer. They have to be detached from the surface before they get sub cultured. Growth limited to surface area. 6

7. Suspension cell Cells which do not require attachment for growth or do not attach to the surface of the culture vessels are anchorage independent cells/suspension cells. All suspension cultures are derived from cells of the blood system because these cells are also suspended in plasma in vitro i.e. They are free floating .,Cells in blood stream . e.g. lymphocytes. Growth is limited to concentration of cells. 7

8. SECONDARY CELL CULTURE: When a primary culture is sub-cultured, it becomes known as secondary culture or cell line. Subculture (or  passage ) refers to the transfer of cells from one culture vessel to another culture vessel. This is periodically required to provide fresh nutrients and growing space for continuously growing cell lines. The process involves removing the growth media and disassociating the adhered cells (usually enzymatically ). Such cultures may be called secondary cultures. 8

9. CELL LINE : A cell line or cell strain may be finite or continuous depending upon whether it has limited culture life span or it is immortal in culture. On the basis of the life span of culture, the cell lines are categorized into two types: Finite cell lines   - The cell lines which have a limited life span and go through a limited number of cell generations (usually 20-80 population doublings) are known as finite cell lines. These cell lines exhibit the property of contact inhibition, density limitation and anchorage dependence. The growth rate is slow and doubling time is around 24-96 hours. 9

10. CONTINUOUS CELL LINES: Cell lines transformed under laboratory conditions or  in vitro  culture conditions give rise to continuous cell lines. These cell lines show the property of  ploidy  ( aneupliody  or  heteroploidy ), absence of contact inhibition (cells growing into a single layer) and anchorage dependence. They grow either in a monolayer or in suspension (see below). The growth rate is rapid and  doubling time  can be 12-24 hours.  10

11. Monolayer cultures   - When the bottom of the culture vessel is covered with a continuous layer of cells, usually one cell in thickness, they are referred to as monolayer cultures. Suspension cultures   - Majority of continuous cell lines grow as monolayers . Some of the cells which are non-adhesive e.g. cells of leukemia or certain cells which can be mechanically kept in suspension, can be propagated in suspension. 11

12. Reference: https://moodle.kent.ac.uk/external/mod/book/view.php?id=2604&chapterid=160 www.slideshare .com 12