The dissociation and separation of bovine adenohypophysial cells

Hirsch, Michael; Lipner, Harry; Leif, Robert C.

doi:10.1007/bf02785059

Cited by 5 publications

(1 citation statement)

References 25 publications

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“…Discontinuous density gradients of Percoll (1 1) have also been used to separate rat somatotropes. This method has also been used to enrich gonadotropes, thyrotropes and lactotropes from bovine anterior pituitary (12) and gonadotrope cells from African catfish (13). More recently, linear Percoll gradients have been used to enrich rat prolactin (PRL) cells (14, 15) and somatotrope cells (16).…”

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confidence: 99%

Application of a Percoll Density Gradient to Separate and Enrich Porcine Pituitary Cell Types

Torronteras

Castaño

Ruíz-Navarro

et al. 1993

J Neuroendocrinology

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The application of a Percoll density gradient cell separation procedure to the pituitary gland of neonatal, prepubertal and mature pigs is described. After enzymatic dispersion, cell viability was 90% to 98% as determined by uptake of Trypan Blue. Recovery of dissociated cells after application of the gradient ranged from 80% to 95%. The dissociated cells were separated in several fractions that were characterized immunocytochemically using different antisera. We obtained highly enriched fractions of gonadotrope (gonadotropins), somatotrope (growth hormone) and lactotrope (prolactin) cells for each age. Concentration of the cell types (purity) was higher than 60% in the following fractions: 1) gonadotropin cells, fraction 15 (1.033 g/cc of density) from mature animals; 2) growth hormone cells, fraction 3 (1.121 g/cc of density) from neonatal animals and fraction 9 (1.087 g/cc) from prepubertal animals; and 3) prolactin cells, fraction 7 (1.094 g/cc) and 10 (1.082 g/cc) from neonatal animals and fraction 14 (1.051 g/cc) from mature pigs. In thyrotrope (thyroid-stimulating hormone) and corticotrope (adrenocorticotropin) cells, enriched fractions were also obtained, although the values of purity were lower (20% to 58%). In conclusion, the proposed cell separation and enrichment technique is suitable for the isolation, purification and examination of porcine pituitary cell types and subpopulations, and offers major advantages such as simplicity, rapidity, efficiency and reproducible results.

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confidence: 99%

Application of a Percoll Density Gradient to Separate and Enrich Porcine Pituitary Cell Types

Torronteras

Castaño

Ruíz-Navarro

et al. 1993

J Neuroendocrinology

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Cell separation methods and selected applications, volumes 1, 2, and 3, edited by Thomas g. Pretlow II and Theresa P. Pretlow. Academic Press, Orlando, 1983, 1984, Vol. 1–330 pp, $37.50; Vol 2–327 pp, $39.50; Vol 3–317 pp, $47.50

Leif

1985

Cytometry

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A short history of the initial application of anti–5‐BrdU to the detection and measurement of S phase

Leif

Stein²,

Zucker

2004

Cytometry Pt A

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Because this article is a bit of history rather than a scientific paper, we will depart from the formality of a journal article. This article has two purposes. The first is to give credit to two deceased individuals, Albert Castro, M.D., and Howard Gratzner, Ph.D. (1). Castro was the key person who was able to produce a polyclonal antibody that was specific for bromodeoxyuridine (BrdU). For 10 years, Gratzner took the lead in converting an idea into a functional assay that affected DNA analysis and led to the development of many other assays with similar methodologies. The second is to inform young scientists that (a) inventions or creations have utility in fields other than the purpose for which they were created; (b) it is possible to do useful work in an institution that certainly would not have been classified as one of the best; and (c) the presence of talented people who are willing to collaborate is invaluable.Scientifically, Gratzner is best known for the development of the first monoclonal antibody (mAb) to BrdU and iododeoxyuridine. This work was the culmination of previous efforts with Robert Leif to develop polyclonal antibodies, which detected BrdU with variable specificity. Collaborations with scientists at the Lawrence Livermore Laboratory encouraged the development of a widely used flow cytometric test for the simultaneous measurements of DNA and BrdU. The use of mAbs to halogenated pyrimidines was a significant scientific advance and currently is an integral tool for the analysis by flow and digital microscopy of the cell cycle.On a personal level, Gratzner's enthusiasm for science and his intelligence, curiosity, loyal interactions with fellow collaborators, and friendship are sorely missed by all who knew him. He was genuinely a warm and nice person who was liked by almost everyone with whom he came in contact (1).Castro was a jovial man who interacted well with his collaborators. He was generous and very knowledgeable. He ran an immunodiagnostics laboratory at the universities of Oregon and Miami and was an expert in the production of antibodies in animals. His warm smile, generous giving of his knowledge, common sense, enthusiasm for scientific research, and ability to get the job done were essential to this project.Because this is a story of events that happened approximately 30 years ago and the existing records are limited, the accuracy of this article is limited by our memory and those of our colleagues who have kindly helped with this project. Of course, this is our version of the events.Cell division and DNA replication are fundamental to biology and specifically to the biology of cancer. This article describes how a simple procedure for the detection and quantitation of DNA synthesis was developed. The precise determination of when S phase occurs in the cell cycle is of use to maximize the selective killing of tumor cells with cycle-specific chemotherapeutic drugs. Of greater significance, studies of the control of the cell cycle, including its detour into apoptosis, can provide extre...

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The dissociation and separation of bovine adenohypophysial cells

Cited by 5 publications

References 25 publications

Application of a Percoll Density Gradient to Separate and Enrich Porcine Pituitary Cell Types

Application of a Percoll Density Gradient to Separate and Enrich Porcine Pituitary Cell Types

Cell separation methods and selected applications, volumes 1, 2, and 3, edited by Thomas g. Pretlow II and Theresa P. Pretlow. Academic Press, Orlando, 1983, 1984, Vol. 1–330 pp, $37.50; Vol 2–327 pp, $39.50; Vol 3–317 pp, $47.50

A short history of the initial application of anti–5‐BrdU to the detection and measurement of S phase

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