Cell culture techniques:primary cells

Question 1

What are the characteristics of the primary cell culture?

Answer 1

Cells derived directly from tissues (unmodified)
Finite lifespan (~6-7 divisions)
They can be grown in 2D or 3D
Cells divide and/or differentiate
Cells carry out normal functions
Good for personalised 
medicine

Question 2

What are the three methods of isolation for a primary cell culture?

Answer 2

1. Cells allowed to migrate out of an explant
2. Mechanical (mincing, sieving, pipetting) or/and enzymatic dissociation (trypsin, collagenase, hyaluronidase, protease, DNAase)

Exception – Haemopoietic cells – Do not need to be disaggregated – They already are

Question 3

Name three techniques which can be used for method of isolation in a primary cell culture ?

Answer 3

• immuno purification
• density configuration
• fluorescence activated cell sorter (FACS)

Question 4

What are the two different categories of primary cell and give examples?

Answer 4

Non-haematopoietic

Haematopoietic

Question 5

Where are the stem cells found in an adult?

Answer 5

Extracted from bone marrow aspirate, fat, mobilised peripheral blood, umbilical cord, embryonal tissue or (or iPSCs!)

Question 6

Name the disadvantages of using a primary cell culture?

Answer 6

Inter-patient variation
Limited
Finite lifespan
Difficult molecular manipulation
Phenotypic instability
Aberrant expression of some genes
Variable contamination