Animal Cell Culture
Primary Explant Culture
Hayflick Limit
Cell Culture Systems
Definition
Steps Overview
A method to culture small pieces of tissue which has been removed surgically from animal tissue or organ (Rao, 2020).
- Acquiring the explant
Primary Cell Culture
Adherent Cell Culture
Suspension Cell Culture
Samples grown and maintained from directly obtained/selected tissue explants OR from individualized cells obtained from the original/parental tissue
Secondary Cell Culture
A concept that aids in the understanding of cellular aging mechanisms.
Monolayer on artificial solid substrate, culture is growing on top of the layer (Thermofisher, n.d.)
Free floating culture in the culture medium (Thermofisher, n.d.)
A regular human cell can divide/grow 40-60 times before reaching its maximum, at which point it will die by apoptosis (programmed cell death).
Cell line
When continuously subcultured, the primary culture can become an established cell line, which can be propagated repeatedly unlike the primary culture and has consistent properties.
Disadvantages
Easy to implement for many cells, easier visual observations, but very limited in terms of growth due to limited surface area (Thermofisher, n.d.)
Require cells that adapts to suspension culture, require regular cell counting, good for bulk or batch harvesting due to its larger volume for cell growth (Thermofisher, n.d.)
Time-consuming
Finite cell lines
Explant-borne contamination
Generation of heterogeneous cells
Continuous cell lines
example
- Cut and clean the explant
- Culturing the explants
Definition
Definition
Advantages & Drawbacks
Advantages & Drawbacks
- Subculturing and constructing cell line
Heterogenous
Advantage: Best culture for In-vivo model/studies and they share the same karyotypes as their parent cells
Three Dimensional
Structure Reconstruction
Subcultures grown from samples produced in the primary cell culture
Dispersed cells are arranged into an organ-like formation to observe the cell-cell and cell-matrix interactions naturally observable.
Low contamination risk
Thermofisher. (n.d.). Adherent cell culture vs. Suspension cell culture. Thermofisher.com. https://www.thermofisher.com/id/en/home/references/gibco-cell-culture-basics/cell-lines/adherent-vs-suspension-culture.html
Definition
Histotypic culture
Advantage: Useful for obtaining large populations of similar cells, able to grow indefinitely (Homogenous, Long lifetime)
Organotypic culture
Co-culturing of cells from various differentiated lineages to induce functionality.
Growing cells in high concentrations similar to that in tissue to observe cell-cell interactions
Disadvantage: Cells may differentiate over a period of time and generate aberrant cells
Disadvantage: High potential for contamination and have limited lifespans
Rao, G. (2020). The What, Why and How of Explant Culture. BiteSizeBio. https://bitesizebio.com/46245/the-what-why-and-how-of-explant-culture/
Definition
Definition
Cell lines with a limited number of cell generations and growth
A set of techniques used to reproducibly cultivate microorganisms at submaximal growth rates at different growth limitations in such a way that the culture conditions remain virtually constant (in ‘steady state’) over extended periods of time
Definition
Definition
Characterization
Anchorage Dependence
Density Limitation
Characterization
Slow Growing
Tumor Like dependency
Immortal like tendency
Fast Growing
Segeritz, C. P., & Vallier, L. (2017). Cell Culture: Growing Cells as Model Systems In Vitro. Basic Science Methods for Clinical Researchers, 151–172. https://doi.org/10.1016/B978-0-12-803077-6.00009-6
Propagation Indefinitely
Propagation until certain generation
Disadvantages
Advantages
Cells have a tendency to differentiate over time in culture
Over time the culture tends to select for aberrant cell
Most cellular characteristic are maintained
Can transform cells to grow indefinitely
Can obtain a large population of similar cells
Disadvantages
Advantages
The more aggressive the cell line, the more it changes over time in culture in cellular level
Not clear how the function of these cells relates to that of other cells (differentiation of healthy and diseased cells)
Easy to obtain large population of cells
Typically easy to manipulate gene expression
Easy to maintain in culture