Garuda - Garba Rujukan Digital

Soni Sopiyana

IRIAP

Author-ID : 2823565

Veterinary

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Primordial germ cells profile incubated for 24 hours in phosphate buffer saline [-] solution Tatan Kostaman; Soni Sopiyana
Jurnal Ilmu Ternak dan Veteriner Vol 22, No 3 (2017): SEPTEMBER 2017
Publisher : Indonesian Center for Animal Research and Development (ICARD)

Gonadal development is a sequential process that can be divided into three major events: the PGCs migration, sex determination and gonadal differentiation. This study was aimed to see the development of PGCs isolated from the gonads of embryos after being incubated for 7 days and then was incubated using a solution of Phosphate Buffer Saline (PBS) [-]. The developing gonad can be isolated from 7 days old chick and can be incubated at a temperature of 37.8oC in a solution of PBS [-]: without Ca2+ and Mg2+. The release of gonadal PGC was observed within 1, 8, 16, and 24 hours after the embryonic gonad was placed in a PBS solution [-]. The results showed that PGCs can be separated from gonadal tissues and can be collected by entering the developing gonad to the PBS [-] solution. The highest percentage of PGCs and survival rate was obtained after gonad was incubated for 1 hour and was not different with 8 hours (P>0.05). Those result was significantly different (P<0.05) with the 16 and 24 hours incubation. The highest purity rate percentage was in the 8 hours incubation, but did not show a significant difference (P>0.05) with the 1 and 16 hours incubation. The percentage of the purity differed (P<0.05) after the 24 hours incubation. It can be concluded that the most appropriate incubation time to obtain PGCs from the KUB chicken embryonic gonad is no more than 8 hours.

Determination of production capacity of circulated primordial germ cells (circulated-PGCs) of KUB chicken using lysis buffer ammonium chloride potassium (ACK) Soni Sopiyana; Iman Supriatna; M. Agus Setiadi; Mohamad Fahrudin
Jurnal Ilmu Ternak dan Veteriner Vol 21, No 1 (2016): MARCH 2016
Publisher : Indonesian Center for Animal Research and Development (ICARD)

In poultry embryos, primordial germ cells (PGCs) are progenitor cells for gametes, which have unique migration pathway. Primordial germ cells arise from epiblast in germinal crescent and circulate through the bloodstream for a short period of time, then leave blood vessel to migrate toward gonads. The aim of this study was to determine the potential production capacity of circulated-PGCs of KUB chicken at different developmental stages of embryo using a rapid and simple method. Seventy five KUB chicken fertile eggs were divided into five groups and incubated at 38.5 0C with a humidity of 60%. Hatching was set to the embryonic development stage of 14-18. The blood was collected through dorsal aorta using micropipette under microscope. The collected blood was placed in a 1.5 ml eppendorf tube which was previously filled with 100 µl phosphate buffered saline without Ca2+ and Mg2+ (PBS-) mixed with fetal bovine serum (FBS) with a ratio of 90%:10%. The PGCs were purified using lysis buffer ammonium chloride potassium method. The results showed that average production of circulated-PGCs per embryo of KUB chicken were significantly affected by stage of embryonic development (P <0.05). The average production of circulated-PGCs at stage 14, 15, 16, 17, and 18 were 37.9; 53.5; 49.8; 38.3; and 33.5 respectively. The number of circulated-PGCs was not different among stages 14, 17 nor 18. The highest number of circulated-PGCs of KUB chicken was obtained at stage 15, so that the isolation and collection of PGCs through the blood circulation was recommended in stage 15.Key Words: KUB Chicken, PGCs, Embryonic Development Stage, Ammonium Chloride Potassium

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