Publisher Summary This chapter focuses on cell culture measurements that can be divided into four major categories: (1) visual methods, which employ the use of light microscopy and devices commonly used in hematology; (2) chemical methods, which employ commonly used analytical biochemical procedures adapted to tissue culture; (3) electronic systems using flow-through cells or apertures for measurement of incorporated dyes or cell numbers; and (4) an array of miscellaneous procedures. The handling of cells prior to analysis is extremely important for a variety of reasons: (1) the exogenous growth media may contain components that are being assayed in the cell, (2) leakage of cellular components may occur if cellular integrity is lost at this point, (3) loss of cells may result from severe mechanical handling, and (4) the use of hydrolytic enzymes may lead to the loss of measurable materials.

Measurement of growth and viability of cells in culture.

Selective induction of DNA synthesis in T and B lymphocytes

Counting actively metabolizing tissue cultured cells.

In proliferating cell populations, the inability to reproduce indefinitely is the only relevant criterion to assess cell lethality. The in vitro colony formation technique (CF) used to determine reproductive death is, however, too slow and has several technical limitations. For finding suitable, more rapid techniques that assessed drug-induced cell killing, a human lymphoma cell line was exposed in vitro to increasing concentrations of adriamycin, bleomycin, and 1,3-bis(2-chloroethyl)-1-nitrosourea for 1 hr. Survival was assayed immediately after treatment and at regular intervals thereafter. Data from CF were compared to those resulting from the following tests: doubling time, labeling index, dye exclusion, 51Cr release, and rate of [3H]thymidine uptake (scintillation index). Dye exclusion and 51Cr release failed to demonstrate any killing effect for the 3 drugs. The percentage of killing calculated from doubling time determinations, although dose dependent, failed to correlate with CF. Scintillation and labeling index values displayed similar temporal fluctuations but were not clearly dose dependent and did not correlate with CF. Thus, CF appears as the most reliable, dose-dependent index of cell lethality. Tests that measure metabolic death grossly overestimate or underestimate killing activity induced by 3 of the most effective antitumor drugs.

https://cancerres.aacrjournals.org/content/canres/36/7_Part_1/2182.full.pdf

Comparison of in vitro methods to determine drug-induced cell lethality.

Publisher Summary The chapter focuses on the developmental aspects of immunity. The ontogenetic development of immune responses in the immature animal and the developmental cellular stages of the immune response in the adult animal are discussed in this chapter. The cellular mechanisms underlying the several immune responses have not received the full attention either to the evolutionary principles underlying the development of all biological systems or to the broad biological rules that govern the proliferative and differentiate activities of cells in any biologically functioning system. The chapter reviews the immunological status of fetal and neonatal mammals as well as several aspects of the ontogeny of the immune response when examined by the most sensitive technique presently available. Developmental stages of immune reactions and their mutual relationships such as relationship of phagocytosis to specific cellular reactions, relationship of delayed hypersensitivity to antibody formation, and dynamics of antibody formation are also presented in this chapter. The increase of immunological capacity with age and the appearance of natural antibodies result solely from specific antigenic stimulation; the chapter discusses the Unitarian concept of cell differentiation and proliferation underlying any form of immunological response. Several methods for studying developmental aspects of immunity and immunological development are also discussed in this chapter.

J.M. Hoskins

Papers

A comparison of methods for estimating the viable count of a suspension of tumour cells.