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Research Development on Cryopreservation Technique to Preserve Avian Semen Kostaman, Tatan; Setioko, A.R.
Indonesian Bulletin of Animal and Veterinary Sciences Vol 21, No 3 (2011)
Publisher : Indonesian Animal Sciences Society

Cryopreservation technique could be used to preserve animal cell, plant or other genetic materials (included semen) in frozen. In this case, the cryopreservation technique is a storageÂ technique that carries out at very low temperature in liquid nitrogen at -196oC. At this temperature, semen does not experience the process of metabolism but still has the ability to live on when used later. Semen that is preserved by cryopreservation technique has unlimited shelf life. This method is more efficient in terms of cost, time, space, and labour than other methods. Cryopreservation techniques can be divided into conventional technique (controlled slow freezing) and rapid freezing technique. Besides cryopreservation of semen, other genetic material from avian that can be cryopreservesed is Primodial Germ Cells (PGC). Balitnak has succesfully isolated the PGC of some Indonesian native chickens. The success of cryopreservation is indicated by not only the high rate of survival, but also the fertility after cryopreservation. Key words: Avian, storage, cryopreservation, semen

Purification Method and Storage of Primordial Germ Cells-Circulation for Preservation of Local Chicken Kostaman, Tatan
Indonesian Bulletin of Animal and Veterinary Sciences Vol 24, No 4 (2014)
Publisher : Indonesian Animal Sciences Society

Primordial germ cells (PGCs) can be used for producing transgenic chickens and preserving genetic material of avian species. Primordial germ cells are precursor of germline cells that could be made proliferation and differentiation to become spermatogonia in testes or oogonia in ovary. Primordial germ cells has a unique migration path, so that the PGCs can be isolated and collected from embryos and propagated and developed through the culture. This paper describes the isolation, collection, purification, storage and transfer of PGCs-circulation of local chickens in Indonesia. Some amount of PGCs-circulation were collected from Gaok chicken at stage 15 as the safe level using cryoprotectant DMSO and some chicks hatched after transfering the PGCs-circulation to recipient embryo. Thus, this technology would be useful for preservation of Indonesian local chicken. Key words: Local chicken, primordial germ cells, cryopreservation

Rate of temperature reduction at cryopreservation primordial germ cells (PGC) of three Indonesian native chicken. Kostaman, Tatan; Sopiyana, S.; Setioko, A.R.
Indonesian Journal of Animal and Veterinary Sciences Vol 16, No 3 (2011)
Publisher : Indonesian Animal Sciences Society

Primordial germ cells (PGCs) are original cells of spermatogonia in the testes or oogonia in the ovary. PGCs in poultry can be harvested and stored in the liquid nitrogen and can be used for conservation as genetic materials of poultry. The objective of this study was to obtain the optimal rate of temperature reduction on PGCs quality from three different Indonesian native chicken after thawing. Fertile eggs obtained from native chicken were incubated for 56 hours to obtain embryo at stage of 14-16. PGCs were isolated from the blood using modified Nycodenz Gradient Centrifugation technique. There after they were kept in a straw and equilibrated for 15 minutes at 5oC and frozen at the rate of temperature reduction of 0.3, 0.5, and 1oC per minute using embryo freezing machine (FHK Fujihara: ET-1). After the temperature reached -30oC, then they were plunged directly into the liquid nitrogen. Recovery rate and viability of PGCs after freezing and thawing were measured. The results of this study showed that the average recovery rate of PGCs that have been frozen at rate of temperature reduction of 1, 0.5, and 0.3oC per minute were 35.6, 43.9, and 44.9% respectively. However the rate of temperature reduction of 0.5 and 0.3oC per minute did not significantly affect the recovery rate. The average percentage of viability of PGCs that were frozen at the rate of 1, 0.5 and 0.3oC per minute were respectively 62.6, 77.5, and 77.4%. It seems that the viability followed the trend of recovery rates where the 1oC per minute reduction was the lowest quality compared to the other two treatments. It is concluded that the reduction of 0.5 or 0.3oC per minute are considered as the ideal temperature reduction when native chicken PGCs are frozen. Key Words: PGCs, Native Chicken, Conservation

Effectiveness of DMSO concentration on recovery rate and viability of primordial germ cell of Gaok chicken Tatan Kostaman; Tuti L. Yusuf; M. Fachrudin; M. A. Setiadi
Jurnal Ilmu Ternak dan Veteriner Vol 22, No 1 (2017): MARCH 2017
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Recent technological developments to produce germ line chimeras with primordial germ cell (PGC) transfer into the recipient embryo provide an opportunity to conserve and retrieval of chicken genetic resources in complete form. The study was conducted to obtain the most effective DMSO percentage to recovery rate and viability of Gaok chicken PGC after freezing which will later be feasible to be transferred. In this study, the eggs of Gaok chicken were incubated for about 2.5 - 3 days to obtain embryos at stages 14 - 16. Blood retrieval was done through the dorsal aorta using micropipettes under microscope. The procedure of PGC isolation of Gaok chicken with centrifugation gradient was using nycodenz as a substance. Commercially available cryoprotectants (dimethyl sulfoxide = DMSO) were used for PGC freezing. Isolated and frozen PGCs of Gaok chicken were diluted with cryoprotectants containing 2.5; 5 and 10% DMSO in fetal bovine serum (FBS). The recovery rate of 2.5; 5 and 10% DMSO concentration were 36.4; 48.2 and 48 % respectively. The viability of PGC after freezing was significantly higher for 5% DMSO compared with 2.5% DMSO (P<0.05), but not different from 10% DMSO. It can be concluded that the concentration DMSO of 5 % was effectve contration in freezing Gaok chicken PGC.

Rate of temperature reduction at cryopreservation primordial germ cells (PGC) of three Indonesian native chicken. Tatan Kostaman; S. Sopiyana; A.R. Setioko
Jurnal Ilmu Ternak dan Veteriner Vol 16, No 3 (2011): SEPTEMBER 2011
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Primordial germ cells (PGCs) are original cells of spermatogonia in the testes or oogonia in the ovary. PGCs in poultry can be harvested and stored in the liquid nitrogen and can be used for conservation as genetic materials of poultry. The objective of this study was to obtain the optimal rate of temperature reduction on PGCs quality from three different Indonesian native chicken after thawing. Fertile eggs obtained from native chicken were incubated for 56 hours to obtain embryo at stage of 14-16. PGCs were isolated from the blood using modified Nycodenz Gradient Centrifugation technique. There after they were kept in a straw and equilibrated for 15 minutes at 5oC and frozen at the rate of temperature reduction of 0.3, 0.5, and 1oC per minute using embryo freezing machine (FHK Fujihara: ET-1). After the temperature reached -30oC, then they were plunged directly into the liquid nitrogen. Recovery rate and viability of PGCs after freezing and thawing were measured. The results of this study showed that the average recovery rate of PGCs that have been frozen at rate of temperature reduction of 1, 0.5, and 0.3oC per minute were 35.6, 43.9, and 44.9% respectively. However the rate of temperature reduction of 0.5 and 0.3oC per minute did not significantly affect the recovery rate. The average percentage of viability of PGCs that were frozen at the rate of 1, 0.5 and 0.3oC per minute were respectively 62.6, 77.5, and 77.4%. It seems that the viability followed the trend of recovery rates where the 1oC per minute reduction was the lowest quality compared to the other two treatments. It is concluded that the reduction of 0.5 or 0.3oC per minute are considered as the ideal temperature reduction when native chicken PGCs are frozen. Key Words: PGCs, Native Chicken, Conservation

Purification Method and Storage of Primordial Germ Cells-Circulation for Preservation of Local Chicken Tatan Kostaman
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 24, No 4 (2014): DECEMBER 2014
Publisher : Indonesian Center for Animal Research and Development

Primordial germ cells (PGCs) can be used for producing transgenic chickens and preserving genetic material of avian species. Primordial germ cells are precursor of germline cells that could be made proliferation and differentiation to become spermatogonia in testes or oogonia in ovary. Primordial germ cells has a unique migration path, so that the PGCs can be isolated and collected from embryos and propagated and developed through the culture. This paper describes the isolation, collection, purification, storage and transfer of PGCs-circulation of local chickens in Indonesia. Some amount of PGCs-circulation were collected from Gaok chicken at stage 15 as the safe level using cryoprotectant DMSO and some chicks hatched after transfering the PGCs-circulation to recipient embryo. Thus, this technology would be useful for preservation of Indonesian local chicken. Key words: Local chicken, primordial germ cells, cryopreservation

Research Development on Cryopreservation Technique to Preserve Avian Semen Tatan Kostaman; A.R. Setioko
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 21, No 3 (2011): SEPTEMBER 2011
Publisher : Indonesian Center for Animal Research and Development

Cryopreservation technique could be used to preserve animal cell, plant or other genetic materials (included semen) in frozen. In this case, the cryopreservation technique is a storage technique that carries out at very low temperature in liquid nitrogen at -196oC. At this temperature, semen does not experience the process of metabolism but still has the ability to live on when used later. Semen that is preserved by cryopreservation technique has unlimited shelf life. This method is more efficient in terms of cost, time, space, and labour than other methods. Cryopreservation techniques can be divided into conventional technique (controlled slow freezing) and rapid freezing technique. Besides cryopreservation of semen, other genetic material from avian that can be cryopreservesed is Primodial Germ Cells (PGC). Balitnak has succesfully isolated the PGC of some Indonesian native chickens. The success of cryopreservation is indicated by not only the high rate of survival, but also the fertility after cryopreservation. Key words: Avian, storage, cryopreservation, semen

Development and Conservation of Gonadal Primordial Germ Cells for Preservation of Local Chicken in Indonesia Tatan Kostaman; S Sopiyana
WARTAZOA, Indonesian Bulletin of Animal and Veterinary Sciences Vol 26, No 3 (2016): SEPTEMBER 2016
Publisher : Indonesian Center for Animal Research and Development

One of the ex situ conservation techniques for poultry that recently developed was to collect primordial germ cell (PGC) or gonadal primordial germ cell (gPGC) that isolated from embryo development. Primordial germ cells (PGC) are embryonic cells that migrate to the gonads and form the precursors of gametes. The unique nature and accessibility of PGC during the early development provides an opportunity to manipulate the poultry germplasm, for example by forming germline chimeras. There are some stages that must be done through isolation and collection of PGC from its resources i.e. blastoderm, embryonic circulation blood and gonad. PGC collection originating from the gonads is one of existing PGC resources and technologies. gonadal PGC have advantages compared with other sources, namely (1) A large number of gonadal PGC can be taken from an embryo; and (2) A collection of gonadal PGC can be used in developing management systems of local avian germplasm conservation. This review is intended to describe the usefulness of isolation and collection technology of gonadal PGC as the local poultry germplasm conservation in Indonesia.

The Conservation on Indonesian Native Chicken Biodiversity Through Primordial Germ Cells Cryopreservation Yuli Arif Tribudi; Tatan Kostaman
Jurnal Ilmu dan Teknologi Peternakan Tropis Vol 9, No 1 (2022): JITRO, Januari 2022
Publisher : Universitas Halu Oleo

ABSTRACT Primordial germ cells (PGCs) can be cryopreserved and used as a potential tool for the preservation of poultry biodiversity. By transplanting donor PGCs into recipient embryos to produce chimera germline, PGCs can migrate to developing gonads where germ cells are produced, thus enabling reproduction of offspring derived from donor PGCs via mating of chimera germline. Collection of PGCs from the circulation of chickens. Fertile eggs were incubated in a portable incubator for 2.5 days. Purification of circulating PGCs with nycodenz. Then the PGCs were frozen using the 5% DMSO cryoprotectant and to determine the success of cryopreservation, the donor PGCs were transplanted into the recipient embryos. The number of circulating PGCs was 50.58 ± 5.79 cells per embryo, the percentage of PGCs viability after thawing was 80.18 ± 1.25%. From the results of the PGCs transplant by looking at the migration of donor PGCs in recipient embryos with PKH-16 staining, it is clear. In conclusion, PGCs cryopreservation in this study can generally support the survival of good chickens and it is hoped that this PGCs conservation protocol can be used for other native chickens.Keywords: PGC, cryopreservation, chicken, conservation

Studi Motilitas dan Daya Hidup Spermatozoa Kambing Boer Pada Pengencer Iris-Sitrat-Fruktosa= Motility and Viability Test of Boer Goat Spermatozoa at Tris-Citrat-Fruktosa Extenders. Tatan Kostaman; I Ketut Suatama
Jurnal Sain Veteriner Vol 24, No 1 (2006): JUNI
Publisher : Fakultas Kedokteran Hewan Universitas Gadjah Mada bekerjasama dengan PB PDHI

Penelitian untuk mengetahui motilitas dan daya hidup spermatozoa kambing Boer pada pengencer trissitrat-fruktosa telah dilaksanakan di Laboratorium Fisiologi Reproduksi, Balai Penelitian Temak, Ciawi-Bogor. Penelitian ini bertujuan untuk mengetahui apakah masih terjadi penurunan kualitas Spermatozoa kambing Boer pada penyimpanan suhu ruangan, karena seperti diketahui semen kambing mudah mengalami kerusakan yang mengakibatkan penurunan kualitas semen, terutama motilitas dan daya hidup spermatozoa. Dua ekor kambing BOW jantan digunakan sebagai sumber semen. Penampungan semen dilakukan sekali dalam seminggu dengan menggunakan vagina buatan. Hasil penelitian menunjukkan bahwa persentase motilitas antara ejakulat 1 dan 2 yang diamati setelah 3 jam pengenceran berbeda nyata (69 vs 67,1%; P0,05). Sementara itu, untuk spermatozoa hidup yang diamati pada 3 dan 6 jam setelah pengenceran tidak berbeda nyata (P>0,05) antara ejakulat 1 dan 2, tetapi tingkat penurunan untuk persentase motilitas dan spermatozoa hidup pada ejakulat 2 memberikan rataan tingkat penurunan yang lebih rendah dibandingkan dengan ejakulat 1 (2,1 vs 4% untuk motilitas dan 0,6 vs 1,8% untuk spermatozoa hidup). Disimpulkan bahwa pengencer Iris-sitrat-fruktosa dapat mempertahankan kualitas semen kambing Boer.

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