Cryopreservation Techniques for Conservation of Tropical Horticultural Species Using Various Explants

2019 ◽  
pp. 579-594 ◽  
Author(s):  
S. K. Malik ◽  
R. Chaudhury
Keyword(s):  
Cryopreservation Techniques

scholarly journals TEKNIK PENYIMPANAN KRIOPRESERVASI PADA BIAKAN AGROBACTERIUM TUMEFACIENS : BAHAN PENELITIAN DI LABORATORIUM TERPADU DAN SENTRA INOVASI TEKNOLOGI UNIVERSITAS JEMBER

2019 ◽  
Vol 2 (2) ◽  
pp. 97-101
Author(s):  
Purnama Okviandari
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Molecular Biology ◽  
Cold Storage ◽  
Cold Temperatures ◽  
Quality Research ◽  
Research Material ◽  
Cooling Devices ◽  
Cryopreservation Techniques ◽  
Storage Technique

Abstrak Teknik Penyimpanan Krioproservasi  Biakan Agrobacterium tumefaciens : Bahan Penelitian  di  Laboratorium Terpadu dan Sentra Inovasi Teknologi Universitas Jember.  Penelitian merupakan kegiatan yang memerlukan bahan yang berkualitas, salah satu cara menjaga kualitas bahan dengan tehnik penyimpanan yang baik. Pemilihan penggunaan tehnik kriopreservasi pada penelitian ini bertujuan menjaga viabilitas  bakteri dalam jangka waktu tertentu. Teknik kriopreservasi yang digunakan  disesuaikan dengan ketersediaan alat pendingin yang ada. Diharapkan dengan penggunaan tehnik kriopreservasi dapat meningkatkan efiisiensi  dan viabilitas sel dalam jangka waktu enam bulan penyimpanan. Penelitian dilakukan di laboratorium Biologi Molekul dan Bioteknologi  menggunakan Agrobacterium tumefaciens strain GV yang sudah terinsersi gen SPS dalam plasmid pKYS (GVpKYS SPS). Bakteri ditumbuhkan pada media yeast, peptone NaCL dengan penambahan antibiotik 100 ppm rifampisin, 12,5 ppm gentamisin dan 50 ppm kanamisin. Dalam biakan bakteri ditambahkan 15% gliserol sebagai kreoprotektan, kemudian dilakukan pembekuan menggunakan nitrogen cair (-196 °C) dan disimpan selama 6 bulan pada suhu dingin. Analisa yang dilakukan adalah uji viabilitas bakteri dan stabilitas genetik diawal dan akhir masa simpan. Hasil penelitian ini diharapkan dapat menyiapkan bahan penelitian yang berkulaitas dan memberikan informasi teknik penyimpanan dingin yang baik dan effisien pada biakan A. tumefaciens khususnya dan bakteri lain pada umumnya.     Abstrac   Cryoproservation Storage Technique for Agrobacterium tumefaciens Culture : Research Material in Center for Devolepment of Advance Science and Technology (CDAST) University of Jember. This research has tole requires quality materials, one of the way to maintain the quality of materials with good storage techniques. Choosing cryopreservation techniques in this study aims to maintain viability of bacteria in a certain period of time. The cryopreservation technique used is adjusted to the availability of existing cooling devices. It is expected that the use of cryopreservation techniques can improve efficiency and viability of cell within six months of storage. The research was conducted in the Molecular Biology and Biotechnology laboratory using Agrobacterium tumefaciens strain GV which was inserted into the SPS gene in plasmid pKYS (GVpKYS SPS). The bacteria are grown on the yeast, NaCL, peptone media. with the addition of 100 ppm antibiotic rifampicin, 12.5 ppm gentamicin and 50 ppm kanamycin. In culture the bacteria added 15% glycerol for cryoprotectant, then it is freezeed using liquid nitrogen (-196 °C) and stored for 6 months in cold temperatures. The analysis carried out was a viability of bacterial test and genetic stability at the beginning and end of the shelf life. The results of this study are expected to be able to prepare quality research materials and provide information on good and efficient cold storage techniques in particular culture of A. tumefaciens and other bacteria.  

scholarly journals Effect of embryo cryopreservation techniques on reproductive and growth traits in rabbits

1999 ◽  
Vol 48 (1) ◽  
pp. 15-24 ◽  
Author(s):  
Josep Cifre ◽  
Manuel Baselga ◽  
Emesto Angel Gómez ◽  
García M. de la Luz
Keyword(s):  
Growth Traits ◽  
Embryo Cryopreservation ◽  
Cryopreservation Techniques

The development of oocyte cryopreservation techniques in blue mussels Mytilus galloprovincialis

2014 ◽  
Vol 80 (6) ◽  
pp. 1257-1267 ◽  
Author(s):  
Hanru Wang ◽  
Xiaoxu Li ◽  
Meiqing Wang ◽  
Steven Clarke ◽  
Mark Gluis
Keyword(s):  
Mytilus Galloprovincialis ◽  
Oocyte Cryopreservation ◽  
Blue Mussels ◽  
Cryopreservation Techniques

Review of ovarian tissue cryopreservation techniques for fertility preservation

2021 ◽  
pp. 102290
Author(s):  
Zahra Bahroudi ◽  
Mahsa Rezaei Zarnaghi ◽  
Melika Izadpanah ◽  
Ali Abedelahi ◽  
Behrooz Niknafs ◽  
...  
Keyword(s):  
Fertility Preservation ◽  
Ovarian Tissue ◽  
Ovarian Tissue Cryopreservation ◽  
Cryopreservation Techniques ◽  
Tissue Cryopreservation

Evaluation of cryopreservation techniques of pancreatic fragments and islets in vitro and in vivo

1988 ◽  
Vol 5 (4) ◽  
pp. 285-293 ◽  
Author(s):  
Kenzo Ishihara ◽  
Hiroshi Taniguchi ◽  
Kazushige Ejiri ◽  
Akimitsu Tsutou ◽  
Keiji Murakami ◽  
...  
Keyword(s):  
Cryopreservation Techniques

56 THE EFFECT OF VITRIFICATION FOR SHEEP OOCYTES VIABILITY

2015 ◽  
Vol 27 (1) ◽  
pp. 121 ◽  
Author(s):  
Y. M. Toishibekov ◽  
R. K. Tursunova ◽  
M. Sh. Yermekova
Keyword(s):  
Polar Body ◽  
Control Group ◽  
Polar Bodies ◽  
Maturation Medium ◽  
Chi Squared ◽  
Fetal Bovine ◽  
Second Polar Body ◽  
Cryopreservation Techniques ◽  
Humidified Air

Advances in reproduction technologies, such as in vitro maturation, IVF, and in vitro culture, stimulated research for efficient cryopreservation techniques for mammalian oocytes. It is well known that the oocyte is the largest cell of an animal's body and as such, is full of water and, in many species, fat, making it difficult to cryopreserve. The objective of this work was to study the effect of vitrification for cryopreservation of the metaphase II plate (MPII) of sheep oocytes. Ovaries from 20 ewes of Kazakh Arkharo-Merino breed were acquired after slaughter and maintained at 37°C in TCM-199. The maturation medium was TCM-199, containing 1 mM of glutamine, 10% FBS, 5 μg mL–1 FSH, 5 μg mL–1 LH, 1 μg mL–1 oestradiol, 0.3 mM sodium pyruvate, and 100 mM cysteamine. The oocytes were incubated in 400 μL of medium in 4-well dishes covered with mineral oil. The IVM conditions were 5% CO2 in humidified air at 39°C for 24 h. Then they were placed for 10 min in a media with Hoechst 33342 (3 μg mL–1) and cytochalasin B (7 μg mL–1) to facilitate the enucleation of the MPII with a minimum volume of ooplasm. The MPII plates were divided into 2 groups: the vitrification group was exposed to vitrification media containing 1.12 M ethylene glycol (ET) + 0.87 M ME2SO for 5 min and was exposed in vitrification media containing 2.24 M ET + 1.75 M ME2SO for 5 min, and then in vitrification solution containing 4.48 M ET + 40% ME2SO + 0.25 M sucrose for 30 s. Oocytes were loaded into cryoloop and plunged into liquid nitrogen (LN2). Oocytes were thawed in a 25°C water bath and then placed in TCM-199 at 20% fetal bovine serum. After 15 min of incubation the oocytes were activated for extrusion of the second polar body in 1 mg mL–1 Ca ionophore for 5 min and washed for 5 min followed by 4 h in 6-DMAP (0.12 mM) + cycloheximide (0.6 μg mL–1). After activation the MPII were washed and cultured for 20 h. The control group received the same treatment, but they were not vitrified. Differences between the experimental groups were tested using Chi-squared test. Our research showed the expulsion of the second polar body after activation was observed in more than 62.2% of the MPII that were not vitrified (control group), whereas 40.5% of vitrified plates had expulsion of polar bodies (P < 0.05). These preliminary studies showed that it is possible to vitrify MPII plates. On the other hand, the drastic reduction of the volume of the sheep oocytes might make cryopreservation possible with greater efficiency.

105. TRANSPLANTED GERM CELLS CAN COLONIZE THE GONADS OF SEXUALLY COMPETENT FISH AND PRODUCE FUNCTIONAL GAMETES

2009 ◽  
Vol 21 (9) ◽  
pp. 24
Author(s):  
S. K. Majhi ◽  
R. S. Hattori ◽  
C. A. Strussmann
Keyword(s):  
Donor Cell ◽  
Cell Behavior ◽  
Cortical Region ◽  
Genital Papilla ◽  
Fluorescent Cell ◽  
Odontesthes Bonariensis ◽  
Sophisticated Equipment ◽  
Elevated Water ◽  
Cryopreservation Techniques ◽  
The Cost

Germ cell (GC) transplantation (GCT) has great potential for seed production and conservation of valuable germlines. Currently available approaches to GCT in fish rely on sophisticated equipment and skills for cell transplantation into the blastodisc of embryos and/or the peritoneal cavity of small, sometimes few millimeters long larvae. Moreover, transplanted individuals may take years to grow to maturity, adding to the cost of producing surrogate gametes. In this context, the use of intragonadal GCT into sexually competent recipients that have been experimentally depleted of endogenous GCs might overcome these constraints. Here we demonstrate the feasibility of xenogeneic GCT in sexually competent fish. Spermatogonial cells isolated from pejerrey (Odontesthes bonariensis, Atherinopsidae) donors were implanted surgically into the testes of congeneric Patagonian pejerrey (O. hatcheri) that were severely depleted of endogenous GCs by treatment with Busulfan (40 mg/kg) and elevated water temperature (25°C) (Fig. 1).Donor cell behavior inside the recipient gonads was tracked using fluorescent cell linkers (CFDA-SE and PKH-26) and showed that transplanted spermatogonial cells were able to migrate towards, settle and multiply at the blind ends (cortical region) of the seminiferous lobules. The presence of donor-derived sperm was confirmed by PCR in 20% of the surrogate Patagonian pejerrey fathers at 6 months and fertilization of pejerrey eggs with surrogate sperm produced 1.2-13.3% pure pejerrey offspring (Fig. 2). These findings indicate that transplantation of spermatogonial cells into sexually competent fish can shorten considerably the production time of surrogate gametes and offspring. Ongoing studies are examining (low-tech) refinements in the proposed approach, such as non-surgical transplantation of GCs through genital papilla, and the suitability of GCT for generation of female gametes, for which cryopreservation techniques have not yet been developed. The results obtained so far have been encouraging and these developments will make GCT invaluable for the timely rescue of fish speciesfacing imminent extinction.

Effects of two cryopreservation techniques on viability and pathogenicity of entomophthoralean fungi

2001 ◽  
Vol 79 (7) ◽  
pp. 861-864 ◽  
Author(s):  
Claudia C. López Lastra ◽  
Ann E. Hajek ◽  
Richard A. Humber
Keyword(s):  
Cryopreservation Techniques

In vitro comparisons of two cryopreservation techniques for equine embryos: Slow-cooling and open pulled straw (OPS) vitrification

2005 ◽  
Vol 64 (7) ◽  
pp. 1619-1632 ◽  
Author(s):  
M. Moussa ◽  
I. Bersinger ◽  
P. Doligez ◽  
F. Guignot ◽  
G. Duchamp ◽  
...  
Keyword(s):  
Slow Cooling ◽  
Cryopreservation Techniques
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