Animal Cell Culture Techniques

Biotechnology & Genetic Engineering Lecture ( 4 ) Assi. Prof. Rajaa Al -Anbaqi



Animal Cell Culture Techniques


Cell culture system involves the cells of particular organ of animal that has been grown in artificial media


Types of cell culture

Many types of cells undergo only a few divisions in vitro before dying out, whereas others will survive for up to a hundred cell generations and some can be propagated indefinitely. These differences, the nature of which are not fully understood, give us four main types of cultured cells:-


a) Primary cell culture:

When cultures are established initially from tissue taken directly from animals, they contain several cell types, most of which are capable of only 5-10 divisions. Due to high cost, inconvenience of getting fresh tissue each time and variation from batch to batch, it is not suitable for use in routine diagnostic work or vaccine production. Furthermore, the donor animals may harbour the latent viruses which can confuse diagnosis or contaminate vaccines. But the primary cultures are very sensitive to many human and veterinary viruses and still used for primary isolation of these viruses.


b) Secondary cell culture:

When primary cells are again passaged in vitro, it is called secondary cell culture.


c) Diploid cell strains:

These are cells that are capable of undergoing a number of divisions in culture that is routinely related to the life span of the species of animals- about 50 for fetal human cells and about 10 for fetal cells from horses and cows. Diploid cell strains of fibroblasts established from human fetuses or embryos are widely used in human diagnostic virology and vaccine production, but diploid strains have not been much used in veterinary vaccine production.

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d) Continuous cell line:


These are cells of a single type that are capable of indefinite propagation in vitro. Such immortal cell lines originate from cancers or by spontaneous transformation of a diploid cell strain. They do not bear the close resemblance to their cell of origin as they undergo many mutations during their prolonged culture. The cells usually have lost the specialized morphology and biochemical abilities. Cells of continuous cell lines are often aneuploid in chromosome number, especially if of malignant origin. Continuous cell lines derived from monkey (MA 104, Vero), dog (MDCK), cattle (MDBK), pig (PK15), Cat (CRFK), mouse (L929, 3T3, hamster (BHK21), rabbit (RK-13), and others are widely used in experimental and diagnostic virology. The great advantage of continuous cell lines over primary cell cultures is that they can be propagated indefinitely by subculturing the cells at regular intervals. Like other cells, they can be preserved for many years when frozen in serum containing medium with added glycerol or DMSO and stored at –80°C or –196°C (liquid nitrogen).