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What is tissue culture?
Tissue culture = organ culture + cell cultureOrgan culture – three dimensional culture of
undisaggregated tissue retaining some or all histological features of tissue in vivo.
Cell culture – derived from dispersed cells taken from original tissue by enzymatic, mechanical or chemical disaggregation.
Historical background (table 1.1)
Devised in 1907 – by Harrison - undisaggregation of cells
Disaggregation of cells and subsequent plating – Rous and Jones, 1916
L929 – first cloned cell strain – Sanford et al., 1948
HeLa – first continuous human cell line – Gey et al., 1952
Sources of tissues
Frog tissueEmbryonated hen’s egg/chicken embryoRodent tissueHuman cells/tissues – HeLa cellsInsect cell lines-Baculovirus
Advantages of Tissue Culture
Consistency and reproducibility of results Reduced statistical analysis of varianceControl physico-chemical environment Control of physiological conditionsChange of selective mediaCryopreservation
Advantages of Tissue Culture
Characterization by immunostainingControl of contaminationReduced volumes of reagentsControl of dose, concentration and timeMicrotitration and roboticsLess moral and ethical issues
Limitations of tissue culture
Chemical and bacterial contaminationStringent aseptic proceduresDisposal of wastesQuantity and cost – medium, pipets etcGenetic instabilityPhenotypic instability - Dedifferentiation
Limitations of tissue culture
Identification of cell type is difficult
- Markers are not always expressed
- Histology is difficult to recreate
In Vitro versus In Vivo cultures
Three-dimensional geometry is lostCultured on substratesSpecific cell interactions are lostCells spread out – proliferateLack systemic components involved in
regulation of homeostasisEnergy metabolism - Glycolysis
Types of Tissue Culture (Organ)
Embryonic organs, adult tissue fragments
High effortCharacterization is
easyHistology is
informativeBiochemical
differentiation
Explant culture
Tissue fragmentsModerate effortCytology and markersHistology is difficultBiochemical
differentiation is heterogeneous
Explant culture
Propagation possible from outgrowth
High intersample variation
Quantitation is difficult
Cell Culture
Disaggregated tissue or primary cell lineLess effortBiochemical, molecular, immunological and
cytological assaysNo histologyBiochemical differentiation is lost
Propagation - Monolayer
Monolayer – Cells will attach to substrateAnchorage dependence – serum derived
glycoproteins, conditioning factors secreted by cells, and cell surface glycoproteins
Propagation - Suspension culture
Survive and proliferate without attachmentAnchorage independentHematopoietic cells, transformed cell lines
and cells from malignant tumors
Summary
Historical backgroundAdvantages and limitationsTypes of tissue culturePropagation of cell linesHistotypic and Organotypic cultureExamination questions
Grant awarded program
This project is funded by a grant awarded under the President’s Community Based Job Training Grant as implemented by the U.S. Department of Labor’s Employment and Training Administration (CB-15-162-06-60). NCC is an equal opportunity employer and does not discriminate on the following basis:
against any individual in the United States, on the basis of race, color, religion, sex, national origin, age disability, political affiliation or belief; and
against any beneficiary of programs financially assisted under Title I of the Workforce Investment Act of 1998 (WIA), on the basis of the beneficiary’s citizenship/status as a lawfully admitted immigrant authorized to work in the United States, or his or her participation in any WIA Title I-financially assisted program or activity.
Disclaimer
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