In vitro culture of growing mouse oocytes

Baran, Madelyn M.; Bachvarova, Rosemary F.

doi:10.1002/jez.1402020219

Cited by 11 publications

(2 citation statements)

References 8 publications

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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

Reproduction

211

108

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Section: Introductionmentioning

confidence: 99%

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

Jutte¹,

Grootegoed²,

Rommerts³

et al. 1981

Reproduction

211

108

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“…Mammalian spermatogenic cells will not develop in MEM containing the same supplements [Steinberger, 19751. Interestingly, oocytes from Xenopus also grow and mature in vitro without follicle cells [Eppig and Dumont, 1976;Wallace and Misulovin, 19781, whereas oocytes from mammals are dependent upon follicle cells for in vitro growth and maintenance [Baran and Bachvarova, 1977;Eppig, 19771. In contrast to other spermatogenic stages, the development of spermatogonia to meiotic prophase and the morphogenesis of elongate spermatids into sperm may require Sertoli cell-specific secretory products since these proesses did not occur in culture. The morphogenesis of late spermatids into sperm may also depend upon mechanical forces established by Sertoli cells.…”

Section: Discussionmentioning

confidence: 99%

Spermatogenic cell differentiation in vitro

Risley

1983

Gamete Research

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Culture conditions that support the in vitro development of many spermatogenic stages from the frog Xenopus laevis are described. Spermatogenic cells were dissociated with collagenase and preelongation stages aseptically isolated by density gradient centrifugation in Metrizamide. The cells were then cultured in modified forms of defined nutrient oocyte medium (DNOM). The development of spermatogenic cells was affected significantly by changes in fetal calf serum concentration, cell density, energy sources, and NaCl concentration. Optimum in vitro spermatid development was obtained when spermatogenic cells were cultured at relatively high densities (3–7 × l07 cells/25 cm2) in DNOM modified to contain 10% heat‐inactivated, dialyzed fetal calf serum, 2 mM 1‐glutamine, 0.1 % glucose, 15 mM HEPES buffer (pH 7.4), and 38.3–48.3 mM NaCl. These culture conditions also supported the differentiation of preelongation spermatids and spermatocytes isolated by density‐gradient centrifugation in Metrizamide and subsequent unit gravity sedimentation in gradients of bovine serum albumin. Approximately 95 % of such isolated spermatids and spermatocytes continued differentiating in vitro for 14 days at in vivo rates. Phase‐contrast and electron microscopy of the cultured cells demonstrated that in vitro differentiation was morphologically normal between the leptotene and elongate spermatid stages. Autoradiographic studies of preleptotene development demonstrated that spermatogonia proliferated and preleptotene spermatocytes developed to zygotene in 12‐day cultures. The results suggest that many spermatogenic stages in Xenopus can develop independent of Sertoli cells, and demonstrate that spermatogenic cell cultures can now be used for in vitro studies of spermatogenesis.

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A comparison between oocyte growth in coculture with granulosa cells and oocytes with granulosa cell‐oocyte junctional contact maintained in vitro

Eppig

1979

J. Exp. Zool.

173

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Evidence is presented that strongly supports the hypothesis that the junctional association between oocytes and granulosa cells must be maintained to promote oocyte growth and development in vitro and that the coculture of oocytes with granulosa cells is not a sufficient condition for oocyte development. Furthermore, it is shown that incorporation of uridine and leucine by oocytes into TCA-insoluble material is significantly greater in granulosa cell-enclosed oocytes than in cocultured oocytes.

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In vitro culture of growing mouse oocytes

Abstract: A method is described for in vitro culture of naked growing mouse oocytes for at least four days. Tests for metabolic function indicate that the oocytes increase in volume, accumulate glucose-6-phosphate dehydrogenase, and maintain a steady level of 3H-uridine and 3H-leucine incorporation.

Cited by 11 publications

References 8 publications

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

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

Spermatogenic cell differentiation in vitro

A comparison between oocyte growth in coculture with granulosa cells and oocytes with granulosa cell‐oocyte junctional contact maintained in vitro

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