174 In vitro spermatogenesis with a novel organ culture system using microfluidic technology

2015 ◽  
Vol 14 (2) ◽  
pp. e174-e174a
Author(s):  
M. Komeya ◽  
T. Yokonishi ◽  
T. Sato ◽  
K. Katagiri ◽  
M. Yao ◽  
...  
Keyword(s):  
Organ Culture ◽  
Culture System ◽  
In Vitro Spermatogenesis ◽  
Organ Culture System ◽  
Microfluidic Technology

P–017 The maintenance of testicular architecture and germ cell in adult testis tissue under organ culture condition based on the gas-liquid interface method

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
M Komeya ◽  
H Odaka ◽  
T Matsumura ◽  
H Yamanaka ◽  
T Sato ◽  
...  
Keyword(s):  
Organ Culture ◽  
Germ Cell ◽  
Germ Cells ◽  
Culture Condition ◽  
Culture System ◽  
Liquid Interface ◽  
Adult Testis ◽  
In Vitro Spermatogenesis ◽  
Organ Culture System

Abstract Study question Can the gas-liquid interface organ culture system that achieved in vitro spermatogenesis in mice also support in vitro spermatogenesis in human adult testis? Summary answer Although the progression of spermatogenesis was not observed, germ cells were maintained without the degeneration of the architecture in both fresh and cryopreserved testicular tissues. What is known already Although the research on in vitro spermatogenesis have been conducted for 100 years, only the organ culture system using gas-liquid interface method achieved in vitro spermatogenesis in mice. It has not been verified whether this culture system can be applied to other mammals including humans and induce spermatogenesis. Study design, size, duration Testicular tissue was obtained from the transgender patients receiving sex reassignment surgery. Testicular specimens were either immediately processed for cultivation or cryopreserved, using a vitrification freezing protocol. Organ culture of testicular fragments was performed in three different media for a maximum period of 3 weeks to evaluate the short-term changes in the cultured tissues (viability, proliferation and maintenance of germ and somatic cells). Participants/materials, setting, methods Fresh and cryopreserved-thawed testis fragments (1–2 mm3) were cultured using the organ culture system in alpha-MEM with knock-out serum replacement (K group), alpha-MEM with lipid-rich BSA (A group) and DMEM with FBS (D group). Luteinizing hormone, follicle stimulating hormone and testosterone were supplemented. The number of germ cells (using DDX4), proliferative activity of germ cells (using EdU assay) and intratubular cell apoptosis (by TdT-mediated dUTP Nick End Labeling) were evaluated by immunohistochemical staining weekly. Main results and the role of chance The architecture of the seminiferous tubules was maintained until the second week of culture in both the fresh and the cryopreserved culture group. The number of DDX4-positive germ cells per seminiferous tubule in groups D, K, and A was 49 ± 24, 55 ± 21, 50 ± 26 cells/tubule in 1 day, 32 ± 13, 42 ± 7, 36 ± 21 cells/tubule in 1week, respectively. The numbers gradually decreased to 26 ± 8, 24 ± 6 and 27 ± 18 cells/tubule, in 2 weeks, respectively, with no difference among the groups. The number of intratubular EdU-positive cells of groups D, K, and A was 0.2 ± 0.2, 2.8 ± 2.1, 1.1 ± 0.8 cells/tubule at 1 day, 0.1 ± 0.2, 0.5 ± 0.6, 0.3 ± 0.6 cells/tubule at 1 week, respectively. The values were 0.01, 0.05, and 0.03 at 2 weeks. Thus, EdU-positive cells drastically decreased from the first week of culture. The number of DDX4-positive germ cells and the intratubular EdU-positive cells in the cryopreserved culture group was not different from that in the fresh culture group. Limitations, reasons for caution Current organ culture systems are incomplete, being unable to induce human in vitro spermatogenesis. Further research is needed to improve culture condition with the aim of producing fertile sperm of infertile adult male patients. Wider implications of the findings: Our organ culture system could maintain testis structure and germ cells. By using the testis tissues of the transgender patients, which are available with their consent, we will promote the investigation of the culture condition necessary for germ cell proliferation and differentiation. Trial registration number Grant-in-Aid for Scientific Research on Innovative Areas 18H05546, Grant-in-Aid for Young Scientists (A) 17H05098 and Takeda Science Foundation

In vivo and in vitro Study of the Effects of Vitamin A Deficiency on Rat Third Molar Development

1986 ◽  
Vol 65 (12) ◽  
pp. 1445-1448 ◽  
Author(s):  
S.S. Harris ◽  
J.M. Navia
Keyword(s):  
Vitamin A ◽  
Organ Culture ◽  
Culture System ◽  
Vitamin A Deficiency ◽  
Third Molar ◽  
In Vitro Study ◽  
Organ Culture System ◽  
Vitamin A Status

We have examined the effect of in vivo vitamin A status on subsequent rat third molar formation and mineralization in an in vitro organ culture system. Vitamin A deficiency imposed during an eight-day in vitro period caused effects very similar to those of vitamin A deficiency imposed on rats in vivo. Analysis of the data also demonstrates that retinoic acid is capable of reversing the interference in mineralization of third molars induced by vitamin A deficiency in the organ culture system.

Usefulness of the organ culture system in the in vitro diagnosis of Celiac Disease: A multicenter study

2006 ◽  
Vol 38 ◽  
pp. S80-S81
Author(s):  
A. Picarelli ◽  
M. Di Tola ◽  
L. Sabbatella ◽  
M.C. Anania ◽  
A. Calabrò ◽  
...  
Keyword(s):  
Celiac Disease ◽  
Organ Culture ◽  
Multicenter Study ◽  
Culture System ◽  
Organ Culture System ◽  
In Vitro Diagnosis

scholarly journals One-step immunochromatographic visual assay for anti-transglutaminase detection in organ culture system: An easy and prompt method to simplify the in vitro diagnosis of celiac disease

2017 ◽  
Vol 32 (1) ◽  
pp. e22195 ◽  
Author(s):  
Marco Di Tola ◽  
Mariacatia Marino ◽  
Rossella Casale ◽  
Raffaele Borghini ◽  
Antonio Tiberti ◽  
...  
Keyword(s):  
Celiac Disease ◽  
Organ Culture ◽  
Culture System ◽  
Organ Culture System ◽  
One Step ◽  
Visual Assay ◽  
In Vitro Diagnosis

Growth hormone and somatostatin directly inhibit gastric ghrelin secretion. An in vitro organ culture system

2007 ◽  
Vol 30 (9) ◽  
pp. RC22-RC25 ◽  
Author(s):  
L. M. Seoane ◽  
O. Al-Massadi ◽  
F. Barreiro ◽  
C. Dieguez ◽  
F.F Casanueva
Keyword(s):  
Growth Hormone ◽  
Organ Culture ◽  
Culture System ◽  
Organ Culture System ◽  
In Vitro Organ Culture ◽  
Ghrelin Secretion

scholarly journals Phenotypic Mapping of The Chicken Embryonic Thymic Microenvironment Developing Within an Organ Culture System

1993 ◽  
Vol 3 (2) ◽  
pp. 147-158 ◽  
Author(s):  
Natalie J. Davidson ◽  
Richard L. Boyd
Keyword(s):  
Organ Culture ◽  
Mhc Class Ii ◽  
Culture System ◽  
Antigen Expression ◽  
Cell Types ◽  
Normal Embryo ◽  
Organ Culture System ◽  
Stromal Component ◽  
Epithelial Antigens

The chicken thymic microenvironment, as it developed in an embryonic thymus organ culture system, was phenotypically mapped using a panel of mAb defining both epithelial and nonepithelial stromal cell antigens. We have previously reported that thymocyte proliferation and differentiation will proceed for up to 6–8 days in thymus organ culture, hence demonstrating the functional integrity of the thymic microenvironmentin vitro. During this time, the stromal component reflected that of the normal embryo with cortical and medullary epithelial areas readily identifiable by both morphology and surface-antigen expression. An abundance of subcapsular and cortical epithelial antigens was detected in the cultured thymus, particularly those normally expressed by the epithelium lining the capsule, trabeculae, and vascular regions (type epithelium) in the adult and embryonic thymus. Medullary epithelial antigens developed in organ culture, although were present in lower frequency than observed in the age-matched embryonic thymus. MHC class II expression by both epithelial and nonepithelial cells was maintained at high levels throughout the culture period. With increasing time in culture, the ratio of epithelial to nonepithelial cells decreased, concurrent with a decrease in thymocyte frequency and suggestive of a bidirectional interaction between these two cell types. Thus, a functionally intact thymic microenvironment appears to be maintained in embryonic thymus organ culture, a model that is currently being exploited to assess the role of stromal antigens, as defined by our mAb, in the process of thymopoiesis.

THE EFFECT OF OESTROGENS ON THE RESPONSE OF BONE TO PARATHYROID HORMONE IN VITRO

1972 ◽  
Vol 54 (1) ◽  
pp. 107-117 ◽  
Author(s):  
D. ATKINS ◽  
JOAN M. ZANELLI ◽  
M. PEACOCK ◽  
B. E. C. NORDIN
Keyword(s):  
Parathyroid Hormone ◽  
Citric Acid ◽  
Organ Culture ◽  
Culture System ◽  
Glucose Consumption ◽  
Organ Culture System ◽  
Mouse Calvaria ◽  
Ethinyl Oestradiol

SUMMARY An organ culture system was used to examine the effects of oestrogens on the response of 5-day-old mouse calvaria to parathyroid hormone (PTH). Parathyroid hormone released calcium and phosphate from the bone and this was associated with an increase in glucose consumption, an accumulation of citric acid and an inhibition of citrate oxidation. Oestradiol, oestriol, oestrone and ethinyl oestradiol all inhibited the PTH-induced release of calcium. The accumulation of citrate was prevented without the PTH-induced block on citrate oxidation being removed, and this was explained in terms of a reduction in glycolysis. Oestradiol, oestriol and oestrone appeared to be of equal potency. However, ethinyl oestradiol was active at much lower doses but appeared to be toxic at higher levels.

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