Anna Steinberger

Mammalian testes in organ culture

Abstract Testes of 14-day-old rats were grown as organ cultures for periods of up to 6 months. The effect of various culture media, gas phases, pHs and incubation temperatures was investigated. Tubular structure with healthy Sertoli cells and mitoses was well maintained under various culture conditions, including growth in chemically defined medium. Spermatocytes survived approximately four weeks without differentiating into spermatids. A cell type, resembling primitive germ cells of younger animals, consistently appeared in seminiferous tubules on the fourth day of culture. Since these cells are not normally observed in the testes of 14-day-old rats, their possible origin in culture is discussed.

In vitro culture of rat testicular cells

Abstract Testicular cells from sexually mature and immature rats were grown in culture under various conditions and were examined by several methods including phase contrast microscopy. The germinal elements did not attach to glass surface and survived for varying periods of time; young spermatids for several days, spermatocytes and possibly spermatogonia for three to four weeks. Mature spermatids were present in the cultures for longer periods. There was no evidence that spermatogenesis. continued in cell cultures. Monolayers formed by non-germinal elements were maintained with subcultures up to seven months. The cells in the monolayers were fibroblast-like and remained euploid. Their origin is under investigation.

Stimulatory effect of vitamins and glutamine on the differentiation of germ cells in rat testes organ culture grown in chemically defined media

Abstract Testes from 4-day-old rats were grown as organ cultures in completely chemically defined media. Spermatogenesis was initiated and progressed to pachytene stage of the meiotic division in complete absence of pituitary gonadotropins. In the presence of combined vitamins A, E and C, or 4 mM-concentration of glutamine, gonocytes differentiated into pachytene spermatocytes at a slightly slower rate than in vivo. Qualitative and quantitative differences were observed between cultures nourished by various media. The possible role of glutamine as an essential factor in differentiation is discussed.

A Familial Centric Chromosome Fragment

An extra centric chromosome fragment was found in the karyotype of an oligospermic male and his apparently normal mother. Both karyotypes were otherwise normal and the origin of the fragment could not

An electron microscopic study of cultured rat testicular fragments

SummaryFragments of testicular tissue of 26-day-old rats were grown as organ cultures during one to six weeks. Electron microscopic studies showed that these tissues can be maintained in vitro for prolonged periods of time, although the most differentiated elements (Leydig and spermatic cells other than spermatogonia) fail to continue their development and degenerate rather rapidly. The connective tissue structures preserve their usual architecture, but the basal membrane of the tubules appears extremely folded and detached from the epithelium. After four weeks in culture, spermatogonia without differentiating are still present, and among them the presence of a “more primitive” type is noted. Sertoli cells are well preserved and ultrastructurally they present the characteristics of the adult type. The possibility exists that differentiation of these two lines of cells may be achieved in vitro if the factors necessary for their growth and differentiation are recognized and incorporated in the culture system.

Electron microscopy of isolated rat testicular cells grown in vitro

SummarySuspensions of viable testicular cells obtained from two groups of rats (one group treated for ten days with 50 I. U. of serum gonadotropins (HCG) daily; one group not previously treated) were cultured for ten days. Numerous cells adhered to the glass to form a confluent monolayer and remained in good condition after ten days. This monolayer contained two cell types identified by electron microscopy: fibroblastic cells and Leydig cells. The relative proportion by which these two cells contributed to the monolayer was related to the condition of the donor when the culture was initiated. Fibroblastic cells were more abundant when the animal was not previously treated with HCG. However Leydig cells almost exclusively formed the monolayer when cultures were begun with testicular cells of HCG-treated rats. Gonadotropins on the other hand did not seem capable of acting in vitro.