Lecture 3: Cell, Tissue And Organ Culture Flashcards
Tissue culture
“Removal of cells, tissues or organs from animals & plants & their placement in an artificial (in vitro) environment that enables them to grow”
Cell culture - growing individual cells normally of a single cell type
Organ culture - whole or fragments
Uses of tissue culture
- investigation of cell metabolism & physiology in vitro
- testing of drugs/chemicals on cells (avoids in vivo - ‘animal testing’)
- developemnt of vaccines …….
History - milestones
- Ringer (1836-1910) ….
Gey
- HeLa cells
- derived from cervical carcinoma from a patient called Henrietta Lack
- cells are immortal (grow constantly)
- first immortal cell line
Eagle and Ham: Typical mammalian culture medium
AMINO ACIDS:
VITAMINS:
SALTS:
MISCELLANEOUS:
PROTEINS (REQUIRED IN SERUM-FREE, CHEMICALLY DEFINED MEDIA): insulin, specific growth factors (do not grow without)
Types of cell/tissue culture
- primary cell culture
- extended (secondary) culture (passage of cells)
- established (continuous) cell lines (transformed & immortal)
Primary cell culture
- Culture derived directly from tissue/organ
- Best resembles tissue in vivo
- Limited growth potential
- Limited life span
- May be a mixture of cell types
Primary cell culture
- Disaggregation of cells (using mechanical or chemical methods)
- Cells grow out from explant
Disaggregation
CHEMICAL:
- EDTA/EGTA (Ca chelators) remove calcium dependent adhesion by molecules such as cadherins and selectins
…..
Extended culture
Continuous cell lines
- Grow indefinitely (immortal)
- Have less stringent growth requirements (no need for serum supplements)
- Grow in suspension (no need for substrate)
…….
Tissue engineering
- Addition of stem cells
- Tissue repair
- Organ substitutes
Tissue engineering
Stem cells
Are undifferentiated cells that can differentiate into specialised cells & can divide to produce more stem cells.
2 types of Stem cells
- Embryonic stem cells: are isolated from the inner cell mass of blastocysts. In a developing embryo stem cells can differentiate into all the specialised cells ….
