Improvement of Mouse Parthenogenetic Blastocyst as Primary Colony Source of Parthenogenetic Embryonic Stem Cell (pESC) using CHIR99021
Improvement of Mouse Parthenogenetic Blastocyst as Primary Colony Source of Parthenogenetic Embryonic Stem Cell (pESC) using CHIR99021
Dwi Budiono1,5, Ratih Rinendyaputri2, Ariyani Noviantari2, Mokhamad Fahrudin3, Ni Luh Putu Ika Mayasari4, Vista Budiariati1,6, Diah Nugrahani Pristihadi1,3, Noer Muhamad Dliyaul Haq1, Arief Boediono3*
ABSTRACT
Parthenogenetic blastocysts as a source of parthenogenetic embryonic stem cells (pESC) has low level of blastocyst rate and quality. CHIR99021 is a material that can increase the formation and quality of blastocysts in buffalo through inhibition of glycogen synthase kinase 3. The purpose of this study was to increase the blastocyst rate and quality of parthenogenetic embryos as a source of pESC primary colonies. Fertilized embryos as positive control group. Diploid parthenogenetic embryos were divided into three treatment groups: CHIR99021 0 mmol/l, CHIR99021 0.003 mmol/l, and CHIR99021 0.01 mmol/l. The embryonic stem cell (ESC) primary colonies were cultured from blastocyst stage embryos. The results of this study showed that CHIR99021 0.003 mmol/l was able to significantly increase the blastocyst rate of parthenogenetic embryos (p<0.05). The quality of blastocysts produced was also significantly improved (p<0.05). Blastocyst rate and quality decreased by increasing CHIR99021 concentration to 0.01 mmol/l. The pESC primary colonies culture showed that blastocysts cultured in CHIR99021 0.003 mmol/l were able to form a pESC primary colony that was similar (p>0.05) to fertilized blastocysts to form ESC primary colonies. These results were significantly improved (p<0.05) compared to those from blastocysts cultured in CHIR99021 0 mmol/l. The characterization results showed that CHIR99021 0.003 mmol/l showed best pluripotency. The conclusion of this study was that the addition of CHIR99021 0.003 mmol/l is most effective in culture of mice parthenogenetic embryos. This is due to CHIR99021 is able to increase blastocyst rate, blastocyst quality, and formation of pESC colonies with improved quality.
Keywords | Blastocyst, CHIR99021, Embryonic stem cell, Parthenogenetic, Primary colony
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