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

2000 ◽  
Vol 260 (1) ◽  
pp. 85-95 ◽  
Author(s):  
Christophe Staub ◽  
Dominique Hue ◽  
Jean-Claude Nicolle ◽  
Marie-Hélène Perrard-Sapori ◽  
Dominique Segretain ◽  
...  
Keyword(s):  
Spermatogenic Cells ◽  
Meiotic Process

In vitro studies of DNA damage and repair in spermatogenic cells from rats and humans

1998 ◽  
Vol 95 ◽  
pp. 213
Author(s):  
C. Bjørge ◽  
A.-K. Olsen ◽  
R. Wiger ◽  
G. Brunborg ◽  
K. Haug ◽  
...  
Keyword(s):  
Dna Damage ◽  
In Vitro Studies ◽  
Spermatogenic Cells ◽  
Dna Damage And Repair

scholarly journals Comparison of biotechnological culture of hypoxia-conditioned rat mesenchymal stem cells with conventional in vitro culture of normoxia-conditioned rat mesenchymal stem cells for testicular failure therapy with low libido in rats

2019 ◽  
Vol 12 (6) ◽  
pp. 916-924 ◽  
Author(s):  
Erma Safitri ◽  
Mas'ud Hariadi
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Functional Outcome ◽  
Leydig Cells ◽  
Control Group ◽  
Spermatogenic Cells ◽  
Hematopoietic Stem ◽  
T1 And T2 ◽  
Testicular Failure

Aim: Biotechnological culture of hypoxia-conditioned (CH) rat mesenchymal stem cells (rMSC-CH) for testicular failure therapy with low libido improves the functional outcome of the testicle for producing spermatogenic cells and repairs Leydig cells in rats (Rattus norvegicus). Materials and Methods: In the first group (T1), rats with testicular failure and low libido were injected with normoxia-conditioned (CN) rMSCs (21% oxygen); in the second group (T2), rats with testicular failure and low libido were injected with rMSC-CH (1% oxygen); in the negative control group (T–), rats with normal testis were injected with 0.1 mL phosphate-buffered saline (PBS); and in the sham group (TS), rats with testicular failure and low libido were injected with 0.1 mL of PBS. Results: Vascular endothelial growth factor expression, as the homing signal, in the groups T2, T–, T1, and TS was 2.00±0.5%, 2.95±0.4%, 0.33±0.48%, and 0±0%, respectively. The number of cluster of differentiation (CD)34+ and CD45+ cells in the groups T– and TS was <20%, whereas that in T1 and T2 groups was >30% and >80%, respectively, showing the mobilization of hematopoietic stem cells (HSCs). The number of spermatogenic cells (spermatogonia, primary spermatocytes, secondary spermatocytes, and spermatid) decreased significantly (p<0.05) in TS compared with that in T–, T1, and T2, whereas that in T2 did not show a significant (p>0.05) decrease compared to that in T–. The improvement in libido, based on the number of Leydig cells producing the hormone testosterone for libido expression, did not increase in T1, whereas T2 was able to maintain the number of Leydig cells significantly compared to that between TS and T1. Conclusion: rMSC-CH culture for testicular failure with low libido showed improvement in the functional outcome of the testicle and in repairing Leydig cells.

Concanavalin A-induced attachment of spermatogenic cells to Sertoli cells in vitro

1982 ◽  
Vol 139 (2) ◽  
pp. 472-475 ◽  
Author(s):  
J GROOTEGOED ◽  
N JUTTE ◽  
F ROMMERTS ◽  
H VANDERMOLEN ◽  
S OHNO
Keyword(s):  
Concanavalin A ◽  
Sertoli Cells ◽  
Spermatogenic Cells

scholarly journals Spontaneous Mutagenesis Is Enhanced in Apex Heterozygous Mice

2004 ◽  
Vol 24 (18) ◽  
pp. 8145-8153 ◽  
Author(s):  
Jessica Huamani ◽  
C. Alex McMahan ◽  
Damon C. Herbert ◽  
Robert Reddick ◽  
John R. McCarrey ◽  
...  
Keyword(s):  
Base Excision Repair ◽  
Germ Line ◽  
Excision Repair ◽  
Spontaneous Mutant ◽  
Spermatogenic Cells ◽  
Ap Endonuclease ◽  
Repair Activity ◽  
Base Excision ◽  
Mutant Frequency

ABSTRACT Germ line DNA directs the development of the next generation and, as such, is profoundly different from somatic cell DNA. Spermatogenic cells obtained from young adult lacI transgenic mice display a lower spontaneous mutant frequency and greater in vitro base excision repair activity than somatic cells and tissues obtained from the same mice. However, spermatogenic cells from old lacI mice display a 10-fold higher mutant frequency. This increased spontaneous mutant frequency occurs coincidentally with decreased in vitro base excision repair activity for germ cell and testicular extracts that in turn corresponds to a decreased abundance of AP endonuclease. To directly test whether a genetic diminution of AP endonuclease results in increased spontaneous mutant frequencies in spermatogenic cell types, AP endonuclease heterozygous (Apex +/−) knockout mice were crossed with lacI transgenic mice. Spontaneous mutant frequencies were significantly elevated (approximately twofold) for liver and spleen obtained from 3-month-old Apex +/− lacI + mice compared to frequencies from Apex +/+ lacI + littermates and were additionally elevated for somatic tissues from 9-month-old mice. Spermatogenic cells from 9-month-old Apex +/− lacI + mice were significantly elevated twofold compared to levels for 9-month-old Apex +/+ lacI + control mice. These data indicate that diminution of AP endonuclease has a significant effect on spontaneous mutagenesis in somatic and germ line cells.

scholarly journals Generation of flagella by cultured mouse spermatids.

1984 ◽  
Vol 98 (2) ◽  
pp. 619-628 ◽  
Author(s):  
G L Gerton ◽  
C F Millette
Keyword(s):  
Electron Microscopy ◽  
Spermatogenic Cells ◽  
Fibrous Sheath ◽  
Acridine Orange Staining ◽  
Transmission Electron ◽  
Fetal Bovine ◽  
Outer Dense Fibers ◽  
Unit Gravity Sedimentation

During the short-term culturing of mouse spermatogenic cells, flagella were generated by round spermatids previously lacking tails. Unseparated germ cells were obtained by enzymatic treatments and round spermatids (greater than 90% pure) were purified by unit gravity sedimentation. As determined by Nomarski or phase-contrast microscopy, no cells had flagella immediately after isolation; flagella were first clearly detected after 6 1/2 h of culture in Eagle's minimal essential medium containing 10% fetal bovine serum and 6 mM lactate. After 24 h, approximately 20% of round spermatids had formed flagella. Multinucleated round spermatids often formed multiple flagella, the number never exceeding the number of nuclei per symplast. Round spermatids were the only spermatogenic cells capable of tail formation. Flagella elongation was blocked by 1 microM demecolcine, an inhibitor of tubulin polymerization. Indirect immunofluorescence localized tubulin in the flagella. As seen by scanning electron microscopy, flagella developed as early as 2 h after culture and continued to elongate over the next 20 h, reaching lengths of at least 19 micron. Transmission electron microscopy demonstrated that flagella formed in culture resembled flagella from Golgi-phase round spermatids in situ; the flagella consisted of "9+2" axonemes lacking other accessory structures such as outer dense fibers and the fibrous sheath. As determined by acridine orange staining of the developing acrosomes, all spermatids that formed flagella in culture were Golgi-phase spermatids. By these criteria, the structures are indeed true flagella, corresponding in appearance to what others have described for early mammalian spermatid flagella in situ. We believe this is the first substantiated report of limited in vitro differentiation by isolated mammalian spermatids.

373 OOCYTE-ACTIVATING CAPACITY OF BOVINE SPERMATOGENIC CELLS AND ACTIVATION PROTOCOLS FOR INTRACYTOPLASMIC INJECTION OF BOVINE ROUND SPERMATIDS

2007 ◽  
Vol 19 (1) ◽  
pp. 302
Author(s):  
C. Kani ◽  
M. Takenaka ◽  
T. Muneto ◽  
M. Yamamoto ◽  
T. Horiuchi
Keyword(s):  
Germ Line ◽  
Oocyte Activation ◽  
Spermatogenic Cells ◽  
Blastocyst Development ◽  
Chi Square ◽  
Mouse Oocytes ◽  
Intracytoplasmic Injection ◽  
Bovine Oocytes ◽  
Activation Capacity

In vitro spermatogenesis can be applied to generate spermatids or spermatozoa and produce a genetically modified male germ line. Intracytoplasmic injection of the spermatids or spermatozoa is an important technique for effective production of offspring. The objective of this study is to evaluate oocyte-activation capacity of bovine spermatids or spermatozoa and to determine the effective activation treatment for in vitro development of bovine oocytes injected with round spermatids. Cryopreserved testicular spermatogenic cells and cauda epididymal spermatozoa obtained from a 1-year-old Japanese bull were used. In the first experiment, we injected bovine round (ROS) and elongated (ELS) spermatids, or testicular (TES) and cauda epididymal (CES) spermatozoa into mouse oocytes to examine their oocyte-activating capacity. The presence of pronuclei within whole-mounted oocytes was observed 4 h after injection. In the second experiment, we injected similar spermatids and spermatozoa into bovine oocytes without additional activation, and examined cleavage and blastocyst development. In the third experiment, bovine oocytes injected with ROS were activated with 7% ethanol or 5 �M ionomycin for 5 min (1 � Et or 1 � Iono) immediately after injection; some were further activated repeatedly at 3 h after injection (2 � Et or 2 � Iono), and some of these were subjected to 1.9 mM 6-dimethylaminopurine (DMAP) for 3 h after the second activation (2 � Et + DMAP or 2 � Iono + DMAP). Data were analyzed by the chi-square test in all experiments. The vast majority of bovine ROS failed to activate mouse oocytes (activation rate 10%). Activation rates of mouse oocytes injected with bovine ELS, TES, and CES were 61%, 75%, and 91%, respectively. The results suggest that oocyte-activation capacity is acquired during transformation from ROS to ELS. Cleavage and blastocyst rates of bovine oocytes injected with CES (59% and 19%, respectively) were significantly higher (P &lt; 0.05) than the rates obtained with injections of TES (37% and 9%) and ROS (5% and 0%) without additional activation. However, cleavage and blastocyst rates of bovine oocytes injected with ROS in the groups of 2 � Et + DMAP (80% and 19%) and 2 � Iono + DMAP (76% and 19%) were significantly higher (P &lt; 0.05) than those in the groups of 1 � and 2 � Et (37% and 2%, 59% and 4%), 1 � and 2 � Iono (10% and 7%, 22% and 4%), or those receiving a sham injection and activated with 2 � Iono + DMAP (43% and 4%). These results demonstrate that intracytoplasmic injection of ROS with repeated Et or Iono activation followed by DMAP treatment is more efficient than single or double Et or Iono activation.

Mono-(2-ethylhexyl) Phthalate (MEHP) Induces Spermatogenic Cell Apoptosis in Guinea Pig Testes at Prepubertal Stage In Vitro

2004 ◽  
Vol 23 (6) ◽  
pp. 349-355 ◽  
Author(s):  
M. A. Awal ◽  
M. Kurohmaru ◽  
M. Ishii ◽  
B. B. Andriana ◽  
Y. Kanai ◽  
...  
Keyword(s):  
Guinea Pig ◽  
Guinea Pigs ◽  
Corn Oil ◽  
Defined Medium ◽  
Tunel Staining ◽  
Spermatogenic Cells ◽  
Control Groups ◽  
Ethylhexyl Phthalate ◽  
Cell Vacuolization

The effects of mono-(2-ethylhexyl) phthalate (MEHP), an active metabolite of di-(2-ethylhexyl) phthalate (DEHP), on prepubertal guinea pig testes in vitro were investigated. The testes of 35-day-old guinea pigs were surgically excised. They were seeded in a defined medium containing antibiotics and administered MEHP at concentrations of 1, 10, and 100 nmol/ml, respectively. The control groups were administered a similar volume of corn oil vehicle. The tissues were incubated for 3, 6, and 9 h. The specimens were collected at 3, 6, and 9 h after treatment. They were fixed in 4%paraformaldehyde or 5% glutaraldehyde. For quantitation of the apoptotic spermatogenic cells, the terminal dUTP nick end-labeling (TUNEL) staining was performed by light microscopy. Detachment and displacement of spermatogenic cells, thin seminiferous epithelia, and Sertoli cell vacuolization were observed. Maximal testicular damage was recognized at 100 nmol/ml 9 h after MEHP treatment. The percentage (%) of apoptotic spermatogenic cells significantly increased at 3, 6, and 9 h after treatment, compared to the control groups. Because the loss of spermatogenic cells by MEHP treatment varies among species, the present study, using guinea pigs, was designed and conducted to obtain further information.

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