Certain Factors Limiting the Size of the Tissue Culture and the Development of Massive Cultures

Earle, Wilton R.; Bryant, Jay C.; Schilling, Edward L.

doi:10.1111/j.1749-6632.1954.tb45887.x

Cited by 70 publications

(11 citation statements)

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“…The mean and standard deviation of each set of measurements (which included at least 20 and, in some cases, more than 200 cells) is given. (10). With both of these techniques only a small per cent of the cell inoculum yielded colonies, and it is impossible to determine whether this low yield is due to trauma suffered by the majority of the cells, or whether it expresses an intrinsic low multiplication potential of the cells from the particular tissues studied.…”

Section: Non-human Serum (Specifically Porcine)mentioning

confidence: 99%

“…These methods also enlarge the scope of genetic investigations on mammalian somatic cells. The ability to establish clonal lines of mammalian cells rapidly is, of itself, only a small improvement on Earle's capillary technique (10). The real genetic contribution of the plating procedures here described is that it permits screening of huge populations so that the rare mutants can be isolated and developed into new strains.…”

Section: Growth In Pooiad Human Serummentioning

confidence: 99%

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Clonal Growth of Mammalian Cells in Vitro

Puck

Marcus

Cieciura

1956

The Journal of Experimental Medicine

1,023

128

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Two methods for simple and rapid plating of single HeLa cells, human, carcinomatous cells, are described. These result in growth and formation of colonies from each single cell. One of these procedures uses irradiated, non-multiplying "feeder" cells to condition the medium. The second requires more gentle handling of the cells, but otherwise is virtually the same as that used in plating bacteria on semisolid, nutrient media. By extension of these methods, it is possible to isolate single mutant colonies and grow pure clonal stocks of animal cells. These genetically uniform strains are much more homogeneous in their behavior than the parental HeLa cell population. Growth curves obtained from developing colonies are highly reproducible. The most active mutant stocks so far isolated display a generation time of 18 to 20 hours. In pooled human serum HeLa cells assume a highly stretched, ameboid form, with marked motility; whereas growth of the same cells in a variety of non-human sera results in tightly packed, columnar, epithelial-like morphology. The two cell types possess volumes, nuclear cross-sections, plating efficiencies, and generation times which are identical within experimental error, but display widely different cross-sectional areas, suggesting that the basic change occurs in the cell surface. It is conceivable that this change may be related to that which enables the cells of a compact tumor to become invasive. Animal cells subjected to the standard trypsinization procedures which involve mechanical trauma and repeated washings in incomplete media leak large amounts of P and suffer impaired ability to reproduce as isolated cells. Application of the methods described in this paper as a tool for quantitative study of normal mammalian cell growth, physiology, genetics, and biochemistry, and the response of cells to drugs, viruses, high energy radiation, and other agents have been indicated.

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Section: Non-human Serum (Specifically Porcine)mentioning

confidence: 99%

Section: Growth In Pooiad Human Serummentioning

confidence: 99%

Clonal Growth of Mammalian Cells in Vitro

Puck

Marcus

Cieciura

1956

The Journal of Experimental Medicine

1,023

128

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“…At this time, Earle et al . had success with growing single mouse cells into colonies from within capillary tubes with conditioned nutrient media derived from mass cultures of growing cells; although, the plating efficiency for this complex process was only 4% (4). Understanding that the key to the growth of single cells in culture was the production of diffusible factors from neighboring cells, Philip Marcus, a graduate student of Puck, devised a unique design with fellow researcher Leo Szilard from the idea, “Since cells grow with high efficiency when they have many neighbors, you should not let the single cells know they are alone” (3).…”

Section: Enabling Replicative Immortalitymentioning

confidence: 99%

Linking the History of Radiation Biology to the Hallmarks of Cancer

2014

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Hanahan and Weinberg recently updated their conceptual framework of the “Hallmarks of Cancer”. The original article, published in 2000, is among the most highly cited reviews in the field of oncology. The goal of this review is to highlight important discoveries in radiation biology that pertain to the Hallmarks. We identified early studies that exemplified how ionizing radiation affects the hallmarks or how radiation was used experimentally to advance the understanding of key hallmarks. A literature search was performed to obtain relevant primary research, and topics were assigned to a particular hallmark to allow an organized, chronological account of the radiobiological advancements. The hallmarks are reviewed in an order that flows from cellular to microenvironmental effects.

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“…As a routine measure, each bottle was trypsinized at least once a week. (d) In inoculation of new bottles, at least 104 cells were always introduced in order to insure the additional mutual support which cells can provide each other under these conditions (7,8,1).…”

Section: Maintenance Of Cultures In a Healthy Acti~dy Multiplyingmentioning

confidence: 99%

Clonal Growth in Vitro of Human Cells With Fibroblastic Morphology

Puck

Cieciura

Fisher

1957

The Journal of Experimental Medicine

302

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In previous papers (1-3) we have described methods for plating single human cells so that each multiplies in isolation to form a well defined colony which constitutes a clonal population. The procedure is simple and quantitative, and completely comparable to the standard plating procedure used by microbiologists for growth of colonies from single bacterial cells. In the work published thus far we have described application of this method only to cells of epithelioid morphology. The present communication demonstrates that fibroblasfic cells, although requiring more stringent care, can also be plated by this technique. Experiments comparing growth and genetic properties of these two cell types obtained from a variety of human organs are presented and some implications of these results for the problem of differentiation are discussed.Definitions.--The terms, done, plating e fidency, and parental strain are used as previously defined (2, 4, 5). Fibroblastic morphology: Elongated form of a glass-attached cell, which is spindle-shaped or possesses multiple, needle-like extensions of the protoplasm, as shown in Fig. 1 a. Colonies of such cells display rough edges. Epithelioid morphology: Cell form consisting of polygonal bodies whose angles are roughly equal in size, as shown in Fig. 1 b. These cells usually pack together fairly tightly, display less surface area and are less migratory than fibroblastic cells (2). In this paper these two terms refer only to the morphologic appearance of the glass-attached cells, without implications concerning their origin in vivo. Genetic or kereditary character: A character, the potentiality for which is transmitted to all the asexual progeny of the original individual even after many passages in an environment which does not permit development or expression of the character. Not all genetic potentialities are genic; i.e. under exclusive control of specific nuclear genes.

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Certain Factors Limiting the Size of the Tissue Culture and the Development of Massive Cultures

Cited by 70 publications

References 1 publication

Clonal Growth of Mammalian Cells in Vitro

Clonal Growth of Mammalian Cells in Vitro

Linking the History of Radiation Biology to the Hallmarks of Cancer

Clonal Growth in Vitro of Human Cells With Fibroblastic Morphology

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