Emil 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 VIVO SURVIVAL OF SPERMATOZOA IN CERVICAL MUCUS.

Abstract 1. Valuable information concerning timing and character of ovulation can be obtained by correlating alterations of cervical mucus with changes in the basal body temperature during the menstrual cycle. 2. Examination of phase lines formed at the interphase between cervical mucus and seminal fluid in vitro gives some indication of the in vivo behavior of these fluids. 3. The consistent absence of spermatozoa from the cervical mucus one day after intercourse or insemination should arouse suspicion about the mucus-semen incompatibility. 4. The presence of many nonmotile and abnormal spermatozoa in cervical mucus 24 hours or longer after intercourse or insemination, in some patients, suggests the possibility that cervical mucus acts as a filter, which prevents ascent of abnormal spermatozoa up the genital tract. 5. Frequently, in vivo spermatozoa survive in cervical mucus for as long as 5 days after insemination or intercourse, and occasionally up to 7 days. Theoretical implications regarding the effectiveness of the “rhythm system” for contraceptive purposes in some women are left to the reader.

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.

Polycystic ovarian disease: A report of 301 patients

Abstract A group of 301 women with polycystic ovarian disease is presented. The 127 patients, whose diagnoses were confirmed surgically, were divided into those with normal or thickened tunica albuginea for purposes of classification. They were further subdivided on the basis of ovarian size (i.e., normal or enlarged). Presenting symptoms of hirsutism, menstrual dysrhythmia, and infertility were compared in each group. No significant differences were noted except that hirsutism occurred more commonly in those with thickened tunica. The effect of prednisone therapy on each of these symptoms was evaluated. Improvement compared favorably with results of wedge resection reported in a recent review. 12

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.

Study of Spermatogenesis and Steroid Metabolism in Cultures of Mammalian Testes

Publisher Summary This chapter describes organ culture technique that permits in vitro maintenance of mammalian testes explants for extended periods of time. Studies concerned with the initiation of spermatogenesis provide evidence that the transition from primordial germ cells—the gonocytes—to adult type A spermatogonia occurs via primitive type A spermatogonia. These cells appeared in culture of testes from animals of all ages and were shown to be capable of reinitiating normal spermatogenesis when transplanted into testes of adult homologous hosts. Using chemically defined media, it is shown in the chapter that in mammalian testes, initiation and progression of spermatogenesis up to late pachytene spermatocytes proceeds in absence of hormones, but completion of the reduction division and formation of spermatids do not take place. The chapter illustrates the beneficial effect of glutamine and vitamins A, C, and E in vitro on the early stages of spermatogenesis. The chapter discusses the attempts that were made to grow the germinal epithelium elements in cell culture system. These cells do not attach to the surface of the culture flasks but remain suspended in the culture media. The germinal elements survive for various periods of time, but no differentiation of any of the cell types present in the inoculum has been observed.

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.

Isosexual pseudoprecocity in a 6-year-old boy with a testicular interstitial cell adenoma

Clinical, hormonal, and pathological findings in a 6-year-old boy with isosexual pseudoprecocity secondary to an androgen-secreting Leydig cell tumor of the testis have been reported. In vivo the tumor secreted androstenedione, testosterone, and dehydroepiandrosterone; the tumor probably also secreted 17α-hydroxyprogesterone, inasmuch as pregnanetriol was present in the urine of this patient. Tumor secretory activity was autonomous and did not alter in response to either HCG or hydrocortisone. Gross and histologic examination of the tumor was compatible with a tumor of interstitial cell origin. Clinical and biochemical remission followed removal of the tumor.

Relation of In Vitro Metabolism of Steroids in Human Testicular Tissue to Histologic and Clinical Findings

The first definitive evidence for androgen secretion by the human testis was provided by experiments demonstrating the formation of radioactive testosterone and androstenedione in spermatic venous effluent following perfusion of the testis with acetate l4C (l). Lucas et al. (2) isolated testosterone from spermatic vein blood, and subsequently a number of investigators demonstrated conversion of suitable steroidal precursors to androgens by human testicular tissue in vitro (3–7). Deficiency of the side-chain cleaving enzyme in testicular tissue of an older man was shown by Axelrod (5). Car-stensen (3) demonstrated in vitro an increased conversion of 17a-hy-droxypregnenolone to testosterone in the presence of ICSH. He suggested that the side-chain cleaving enzyme may be activated by gonadotropins. A suppression of the activity of the side-chain cleaving enzyme and the 17α-hydroxylase, but no change, or possibly an increase, in the activity of 17 s-hydroxysteroid dehydrogenase was demonstrated in the microsomal fraction of testicular tissue from a patient with prostatic carcinoma treated with estrogens (8). If the assumption is made that estrogen suppressed patients’ endogenous gonadotropins, the decrease in 17–20 lyase may be interpreted as confirmation of Carstensen’s findings.