"A Diffusible Agent of Mouse Sarcoma, Producing Hyperplasia of Sympathetic Ganglia and Hyperneurotization of Viscera in the Chick Embryo," by Rita Levi-Montalcini and Viktor Hamburger, appeared in 1953 in the Journal of Experimental Zoology. The paper provided the first evidence that nerve growth factor is a diffusible substance. Nerve growth promoting tumors were implanted into developing embryos to determine whether the tumors stimulated growth by direct contact or by a diffusible substance. The tumors were implanted into different parts of the embryo; one set of experiments implanted the tumor directly in the embryo, while another set of experiments placed the tumor on an exterior membrane. The membrane provided a barrier to direct contact with the nervous system, and allowed some chemical interaction between the tumor and the embryo. The tumor stimulated growth in both orientations, demonstrating that nerve growth factor was a chemical agent. The paper ends with two possible conclusions for the mechanism of nerve growth, the tumor may have been directly stimulating the ganglia by a diffusible signal, or it may have reduced the resistance of the chick tissues to the nerve growth. The term "nerve growth factor" is not explicitly used in this paper, and is referred to as a "diffusible agent" or a "growth promoting agent."

Nicole Marthe Le Douarin was one of the first progressive female pioneers of developmental and embryological research. Some of her most notable and ground-breaking work involves grafting quail and chicken embryos together in order to study the developmental fate of each contributing embryo. Le Douarin was born in Brittany, France, on 20 August 1930. As an only child she was inspired by her mother, a school teacher at the time, to develop a passion for learning. According to Le Douarin her father was an open-minded businessman who, likely because she was an only child, raised her much like a boy. In 1944 Le Douarin was forced to move out of her hometown of Lorient and attend a boarding school in Nantes, France, to escape the invasion of German forces during World War II. After the war, she returned to her high school in Lorient, where she received her baccalaureate in 1949. During her last year in Lorient, when she was only seventeen, Douarin met her fianc_. After graduating, and against the wishes of her mother, she moved with him to Paris where they attended the Sorbonne. In 1951, after three years of courtship and university classes together, the couple was married. Le Douarin graduated from the Sorbonne with a degree in the natural sciences in 1954. Instead of immediately continuing to graduate school, she chose to teach science at a local high school and raise a family.

Embryonic images displayed in Life magazine during the mid-twentieth century serve as a representation of technological advances and the growing public interest in the stages of embryological development. These black-and-white photographs portray skeletal structures and intact bodies of chicken embryos and human embryos and fetuses obtained from collections belonging to universities and medical institutions.

August Antonius Rauber was an embryologist and anatomist who examined gastrulation in avian embryos. He examined the formation of the blastopore, epiblast, and primitive streak during chick development. Subsequent researchers have further studied Rauber's findings, which has led to new discoveries in embryology and developmental biology.

Gastrulation is an early stage in embryo development in which the blastula reorganizes into three germ layers: the ectoderm, the mesoderm, and the endoderm. Gastrulation occurs after cleavage but before neurulation and organogenesis. Ernst Haeckel coined the term; gaster, meaning stomach in Latin, is the root for gastrulation, as the gut is one of the most unique creations of the gastrula.

A node, or primitive knot, is an enlarged group of cells located in the anterior portion of the primitive streak in a developing gastrula. The node is the site where gastrulation, the formation of the three germ layers, first begins. The node determines and patterns the anterior-posterior axis of the embryo by directing the development of the chordamesoderm. The chordamesoderm is a specific type of mesoderm that will differentiate into the notochord, somites, and neural tube. Those structures will later form the vertebral column. In the chick embryo, the node is referred to as Hensen's node because of its discoverer, Viktor Hensen, who first described the node in 1875. The discovery of Hensen's node has helped to answer questions of axis formation and has allowed experimental embryologists to further investigate vertebrate embryonic development.

In 1951 Viktor Hamburger and Howard Hamilton created an embryonic staging series from a combination of photographs and drawings from other researchers. The Hamburger-Hamilton stages are a sequence of images depicting 46 chronological stages in chick development. The images begin with a fertilized egg and end with a fully developed chick. The Hamburger-Hamilton staging series was produced in order to replace a previous chick staging series created in 1900. The earlier attempt lacked specific details and staged the chick embryo by using only morphological characteristics. As chicks were, and still remain, model organisms for experimental embryology, it was important to create a staging series with descriptions for determining the approximate age of a developing chick embryo.

Conrad Hal Waddington's "Experiments on the Development of Chick and Duck Embryos, Cultivated in vitro," published in 1932 in Philosophical Transactions of the Royal Society of London, Series B, compares the differences in the development of birds and amphibians. Previous experiments focused on the self differentiation of individual tissues in birds, but Waddington wanted to study induction in greater detail. The limit to these studies had been the amount of time an embryo could be successfully cultivated ex vivo. Waddington applied in vitro cell culturing techniques to this experiment, as opposed to the chorio-allantoic technique used in many earlier studies. Culturing in vitro consisted of placing the embryo on a clot of adult chicken blood plasma and chick embryo extract in a watch glass. Experiments reported in this paper were divided into three main sections: the development of the embryos in vitro, induction by the endoderm, and induction by the primitive streak.

German embryologist Viktor Hamburger came to the US in 1932 with a fellowship provided by the Rockefeller Foundation. Hamburger started his research in Frank Rattray Lillie's laboratory at the University of Chicago. His two-year work on the development of the central nervous system (CNS) in chick embryos was crystallized in his 1934 paper, "The Effects of Wing Bud Extirpation on the Development of the Central Nervous System in Chick Embryos," published in The Journal of Experimental Zoology. Hamburger was able to use the microsurgical techniques that he had learned from Hans Spemann to show how wing buds influence the development of the CNS in chick embryos. This paper is one of several among Hamburger's important studies on chick embryos and represents the empirical and theoretical cornerstone for his further research on central-peripheral relations in the development of the nervous system.

In the 1910s, Alexis Carrel, a French surgeon and biologist, concluded that cells are intrinsically immortal. His claim was based on chick-heart tissue cultures in his laboratory that seemed to be able to proliferate forever. Carrel's ideas about cellular immortality convinced his many contemporaries that cells could be maintained indefinitely. In the 1960s, however, Carrel's thesis about cell immortality was put into question by the discovery that human diploid cells can only proliferate for a finite period. As it was gradually recognized that chick cells only have a finite proliferative life span in vitro as well, historians and scientists alike attempted to identify experimental errors that could have led to the extremely long life of Carrel's "immortal" chick-heart tissue cultures. Those reassessments not only point out potential experimental mistakes in pioneer tissue culture work in the early twentieth century, but are also relevant to current discussions about the different life spans of germ line cells, embryonic and adult stem cells, normal somatic cells, and cancer cells.

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