Morphological characteristics of male germ cells of rats in contact with Sertoli cells in vitro

Palombi, Fioretta; Ziparo, Elio; Rommerts, F. F. G.; Grootegoed, J. Anton; Antonini, Marco; Stefanini, Mario

doi:10.1530/jrf.0.0570325

Cited by 43 publications

(15 citation statements)

References 7 publications

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“…However, in their systems, no haploid cells were formed. The use of bicameral chambers in the present work is most probably an important parameter explaining at least partly, the difference between our results and those of others [25][26][27][28]. Indeed, under our conditions, Sertoli cells maintained morphological features of healthy polarized Sertoli cells throughout the culture period.…”

Section: Discussioncontrasting

confidence: 60%

“…Palombi et al [25] and Tres and Kierszenbaum [26] reported cytological studies of culture of fragments of seminiferous epithelium, prepared from 3-week-old rats, showing well-preserved morphological characteristics of many germ cells in contact with Sertoli cells after culture periods up to 9 days. However, in their systems, no haploid cells were formed.…”

Section: Discussionmentioning

confidence: 99%

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The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

Staub

Hue

Nicolle

et al. 2000

Experimental Cell Research

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The aim of the present study was to assess whether the whole meiotic process of spermatogenic cells is able to take place in vitro. Fragments of seminiferous tubules from 20-to 22-or 28-day-old rats were seeded in medium containing 0.2% fetal calf serum in bicameral chambers and then cultured for 4 weeks in a chemically defined medium. The differentiation of meiotic germinal cells was followed by four criteria: (i) ultramicroscopic examination of the different types of germ cells present in the cell layer throughout the culture period; (ii) determination of the changes in DNA content per nucleus of the cell population seeded with time in culture; (iii) assessment of the ability of germinal cells to transcribe genes expressed after completion of meiosis; and (iv) monitoring the fate of BrdU-labeled leptotene spermatocytes. The ultrastructural study showed that the overall organization of the cells in the culture well recalls that of the seminiferous epithelium throughout the culture period. Moreover the identification of young round spermatids 21 days after seeding suggested that these spermatids had been formed very recently in culture. Determination of DNA content per nucleus showed that a 1C cell population could be observed after several days of cultures reaching 6 to 10% of total cells. An exponential-like increase in the amounts of the mRNAs encoding for TP1 or TP2 occurred from the time when 1C cells appeared in the culture until the end of the experiment. Finally, BrdU-labeled leptotene spermatocytes differentiated into pachytene spermatocytes and then into secondary spermatocytes, and BdrU-labeled round spermatids were observed from Day 21 of culture onward. Taken together these results indicate that the whole meiotic process from leptotene spermatocyte to round spermatid can indeed occur in vitro under the present culture conditions.

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Section: Discussioncontrasting

confidence: 60%

Section: Discussionmentioning

confidence: 99%

The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

Staub

Hue

Nicolle

et al. 2000

Experimental Cell Research

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“…In 1979, the Stefanini team confirmed the previous results with a quite different culture system of seminiferous tubules (Palombi et al, 1979). They took their inspiration from successes of the Dorrington group for cell preparation (Dorrington and Armstrong, 1975) and from the Steinberger group (Steinberger and Steinberger, 1966) for the incubation medium.…”

Section: Cultures On Impermeable Supportsmentioning

confidence: 74%

“…These cell interactions seem to improve the survival of germ cells in culture. In such a system, it is possible for germ cells from immature or prepuberal animals to survive 1 month in vitro (Eddy and Kahri, 1976;Palombi et al, 1979), but nothing has been said about their ability to differentiate in this lapse of time. The fact that the tubules are seeded open may still improve the life expectancy of the cultured cells, simply by making the exchange of metabolites between the culture medium and a maximum of cells easier (Toebosch et al, 1989).…”

Section: Cultures On Impermeable Supportsmentioning

confidence: 99%

A Century of Research on Mammalian Male Germ Cell Meiotic Differentiation In Vitro

Staub

2001

Journal of Andrology

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Many germ cell studies over the last century have focused on testicular cell culture (Wolff and Haffen, 1965;Russell and Steinberger, 1989;Kierszenbaum, 1994). The first germ cell culture systems were directed at studying gonadal embryogenesis and male germ cell differentiation. Today's culture systems are powerful tools that bring about new knowledge and applications to the study of spermatogenesis. The culture of Sertoli cells is mentioned only as a coculture process with germ cells. The main focus of this review is the differentiation of male germ cells, and more precisely, the meiotic step of that differentiation.The various studies can be divided into 3 main categories depending on the historical advancement (early attempts at gametogenesis in vitro, intermediate progress, and latest refinements). Examples highlighting the strong and weak points of each period are summarized in this review.

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“…It has been shown that testicular LDH-X catalyses preferentially lactate oxidation and is localized in cytosol and mitochondria (Blanco, Burgos, Gérez de Burgos & Montamat, 1976;. This, as well as the localization of mitochondria in early round spermatids close to the cell surface (Clermont & Rambourg, 1978) (Palombi et al, 1979 (Donahue & Stern, 1968 (Biggers, Whittingham & Donahue, 1967;Zeilmaker & Verhamme, 1974;Hillensjö, Hamberger & Ahrén, 1975;Eppig, 1976), but growing oocytes can survive in the absence of pyruvate when cultured in the presence of follicular granulosa cells (Baran & Bachvarova, 1977;Eppig, 1977;Bachvarova, Baran & Tejblum, 1980 …”

Section: Introductionmentioning

confidence: 99%

Exogenous lactate is essential for metabolic activities in isolated rat spermatocytes and spermatids

Jutte¹,

Grootegoed²,

Rommerts³

et al. 1981

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Morphological characteristics of male germ cells of rats in contact with Sertoli cells in vitro

Cited by 43 publications

References 7 publications

The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

The Whole Meiotic Process Can Occur in Vitro in Untransformed Rat Spermatogenic Cells

A Century of Research on Mammalian Male Germ Cell Meiotic Differentiation In Vitro

Exogenous lactate is essential for metabolic activities in isolated rat spermatocytes and spermatids

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