In Vitro Spermatogenesis Using an Organ Culture Technique

2012 ◽  
pp. 479-488 ◽  
Author(s):  
Tetsuhiro Yokonishi ◽  
Takuya Sato ◽  
Kumiko Katagiri ◽  
Takehiko Ogawa
Keyword(s):  
Organ Culture ◽  
Culture Technique ◽  
In Vitro Spermatogenesis

scholarly journals A Protocol on in Vitro Propagation of Arbuscular Mycorrhizal Fungi using Root Organ Culture Technique

2017 ◽  
Vol 31 (1-2) ◽  
pp. 17-24
Author(s):  
Hari Prasad Aryal
Keyword(s):  
Arbuscular Mycorrhizal Fungi ◽  
Organ Culture ◽  
Mycorrhizal Fungi ◽  
In Vitro Propagation ◽  
Arbuscular Mycorrhizal ◽  
Culture Conditions ◽  
Culture Technique ◽  
The Past ◽  
Root Organ Culture

 The technique of in vitro propagation of Arbuscular mycorrhizal fungi has been developed over the past few decades and opens up areas of studying plant-fungi interactions. It is a scientific break through, especially for the study of the Arbuscular mycorrhizal fungi, since these obligate symbionts depend on host plant. The objective of this paper is to find out the in vitro culture of Arbuscular Mycorrhizal Fungi using Root Organ Culture technique. Ascertain of root colonization of these fungi could be affected in vitro without undertaking complex and complicated culture conditions. This could form an economically viable technique for root organ culture of Arbuscular mycorrhizal fungi.

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

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

scholarly journals Transcriptome analysis reveals inadequate spermatogenesis and immediate radical immune reactions during organ culture in vitro spermatogenesis

2020 ◽  
Vol 530 (4) ◽  
pp. 732-738 ◽  
Author(s):  
Takeru Abe ◽  
Hajime Nishimura ◽  
Takuya Sato ◽  
Harukazu Suzuki ◽  
Takehiko Ogawa ◽  
...  
Keyword(s):  
Organ Culture ◽  
Transcriptome Analysis ◽  
Culture In Vitro ◽  
Immune Reactions ◽  
In Vitro Spermatogenesis

Development and evaluation of a porcine in vitro colon organ culture technique

2016 ◽  
Vol 52 (9) ◽  
pp. 942-952 ◽  
Author(s):  
Matheus O. Costa ◽  
John C. S. Harding ◽  
Janet E. Hill
Keyword(s):  
Organ Culture ◽  
Culture Technique

IN VITRO METHODS FOR THE STUDY OF THE ADENOHYPOPHYSIAL FUNCTIONS TO SECRETE GONADOTROPHIN

1971 ◽  
Vol 68 (1_Suppl) ◽  
pp. S27-S40 ◽  
Author(s):  
T. Kobayashi ◽  
T. Kigawa ◽  
M. Mizuno ◽  
T. Watanabe
Keyword(s):  
Cell Culture ◽  
Organ Culture ◽  
Cultured Cells ◽  
Cell Sheet ◽  
Short Term ◽  
Hypothalamic Extract ◽  
Numerous Cell ◽  
Single Cell Culture ◽  
Almost All

ABSTRACT There are several in vitro methods to analyse the function of the adenohypophysis or the mechanisms of its regulation. The present paper deals with single cell culture, organ culture and short term incubation techniques by which the morphology and gonadotrophin-secreting function of the adenohypophysis were studied. In trypsin-dispersed cell culture, the adenohypophysial cells showed extensive propagation to form numerous cell colonies and finally develop into a confluent monolayer cell sheet covering completely the surface of culture vessels. Almost all of the cultured cells, however, became chromophobic, at least at the end of the first week of cultivation, when gonadotrophin was detectable neither in the culture medium nor in the cells themselves. After the addition of the hypothalamic extract, gonadotrophin became detectable again, and basophilic or PAS-positive granules also reappeared within the cells, suggesting that the gonadotrophs were stimulated by the extract to produce gonadotrophin. In organ culture and short term incubation, the incorporation of [3H] leucine into the adenohypophysial cells in relation to the addition of hypothalamic extract was examined. It was obvious that the ability to incorporate [3H] leucine into the gonadotrophs in vitro was highly dependent upon the presence of the hypothalamic extract.

scholarly journals Nano-Elicitation as an Effective and Emerging Strategy for In Vitro Production of Industrially Important Flavonoids

2021 ◽  
Vol 11 (4) ◽  
pp. 1694
Author(s):  
Amna Komal Khan ◽  
Sidra Kousar ◽  
Duangjai Tungmunnithum ◽  
Christophe Hano ◽  
Bilal Haider Abbasi ◽  
...  
Keyword(s):  
World Market ◽  
Culture Technique ◽  
In Vitro Cultures ◽  
Defense System ◽  
In Vitro Production ◽  
Metallic Oxide ◽  
Global Demand ◽  
Over Exploitation ◽  
Vitro Production

Flavonoids represent a popular class of industrially important bioactive compounds. They possess valuable health-benefiting and disease preventing properties, and therefore they are an important component of the pharmaceutical, nutraceutical, cosmetical and medicinal industries. Moreover, flavonoids possess significant antiallergic, antihepatotoxic, anti-inflammatory, antioxidant, antitumor, antiviral, and antibacterial as well as cardio-protective activities. Due to these properties, there is a rise in global demand for flavonoids, forming a significant part of the world market. However, obtaining flavonoids directly from plants has some limitations, such as low quantity, poor extraction, over-exploitation, time consuming process and loss of flora. Henceforth, there is a shift towards the in vitro production of flavonoids using the plant tissue culture technique to achieve better yields in less time. In order to achieve the productivity of flavonoids at an industrially competitive level, elicitation is a useful tool. The elicitation of in vitro cultures induces stressful conditions to plants, activates the plant defense system and enhances the accumulation of secondary metabolites in higher quantities. In this regard, nanoparticles (NPs) have emerged as novel and effective elicitors for enhancing the in vitro production of industrially important flavonoids. Different classes of NPs, including metallic NPs (silver and copper), metallic oxide NPs (copper oxide, iron oxide, zinc oxide, silicon dioxide) and carbon nanotubes, are widely reported as nano-elicitors of flavonoids discussed herein. Lastly, the mechanisms of NPs as well as knowledge gaps in the area of the nano-elicitation of flavonoids have been highlighted in this review.

Studies of Metabolism Mediated Mutagenicity In Vitro

1990 ◽  
Vol 18 (1_part_1) ◽  
pp. 243-250
Author(s):  
Dag Jenssen ◽  
Lennart Romert
Keyword(s):  
Cell Lines ◽  
Biological Effects ◽  
Animal Experiments ◽  
Culture Technique ◽  
Organ Perfusion ◽  
Isolated Cells ◽  
Toxicological Effect ◽  
In Vitro Methods

To understand the cause of the biological effects of xenobiotic metabolism in mammals, investigators have traditionally performed animal experiments by comparing the results of biochemical methods, such as measurement of enzyme activity analysis of the metabolites produced, with the observed toxicological effect. This article deals with in vitro methods for genotoxicity combined with drug metabolising preparations at the organelle, cell or organ levels, as exemplified by microsome preparations, isolated cells/cell lines and organ perfusion systems, respectively. The advantage of some of these methods for studying metabolism-mediated mutagenicity is that the measured endpoint reflects not only the bioactivating phase I reactions, but also the detoxifying phase II reactions, and the transfer of the non-conjugated reactive metabolites to other cells and their ability to cause mutations in these cells. In vivo, all these events are important factors in the initiation of cancer. A mechanistic advantage of the methods for metabolism-mediated mutagenicity in vitro is that the relevance of the different steps in metabolism for the mutational events can seldom be investigated in an in vivo assay. Furthermore, human studies can easily be performed using the co-culture technique with isolated human cells or cell lines.

scholarly journals Insights into in vitro spermatogenesis in mammals: Past, present, future

2017 ◽  
Vol 84 (7) ◽  
pp. 560-575 ◽  
Author(s):  
Amir Fattahi ◽  
Zeinab Latifi ◽  
Tohid Ghasemnejad ◽  
Hamid Reza Nejabati ◽  
Mohammad Nouri
Keyword(s):  
In Vitro Spermatogenesis
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