Introduction Chapter 1. What is tissue culture? Tissue culture = organ culture + cell culture Organ culture – three dimensional culture of undisaggregated.

Introduction

Chapter 1

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

Various tissue culture applications

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

Organ culture

No propagationHigh intersample

variationQuantitation is

difficult

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

Cell Culture

Propagation is possibleLow intersample variationHigh studies of quantitation

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

Cell Culture

Histotypic culture/Histoculture

Tissue culture of one cell typeHigh density

Organotypic culture

Restricted to organ cultureMore than one cell interaction

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.

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