Produksi Embryonic Stem Cell (Esc) Line dari Blastosis Mencit dengan Metode Immunosurgery

jbmi managerxot; Ratih Rinendyaputri; Nike Susanti

doi:10.22435/jbmi.v3i1.1654

jbmi managerxot
Ratih Rinendyaputri
Nike Susanti

DOI: https://doi.org/10.22435/jbmi.v3i1.1654

Keywords: Blastosis, Inner cell mass, ICM, ESC

Abstract

Abstract
Embryonic Stem Cells (ESC) are pluripotent stem cells which has the ablility to self renew and differentiate into form all cells in the body. Inner Cell Mass (ICM) as a source of ESC that can be obtained from blastocyst stage of embryos. Isolation can be done by several methods such as mechanical, enzymatic and immunosurgery. This study aimed to observe the effectiveness of usage immunosurgery method to obtain the ICM from the blastocyst stage of embryo. Blastocyst stage of embryos were obtained from a strain of Swiss Webster female mice that had been stimulated using pregnant mare 's gonadotropin (PMSG) and human chorionic gonadotropin (hCG). Inner Cell Mass (ICM) were cultured and observed of the attachment level (attachment rate/AR), the rate of primary colony formation (primary colony/PC) and the morphology of ESC line. The results showed that the ICM were isolated using methods immunosurgery have AR and PC respectively 78,57% and 71,00%. Embryonic Stem Cell (ESC) can differentiate after several passages by forming embryoid body (EB). This study shows that immunosurgery is an effective method for producing ESC line from mice embryos of blatocyst stage.

Key words: Blastocyst, Inner cell mass, ICM, ESC

Abstrak
Embryonic Stem Cell (ESC) merupakan sumber stem cell yang bersifat pluripoten, yaitu sel yang mampu membelah dan berdiferensiasi menjadi semua tipe sel di dalam tubuh. Inner Cell Mass (ICM) sebagai sumber ESC dapat diperoleh dari embrio tahap blastosis. Isolasi ICM dapat dilakukan dengan beberapa metode seperti mekanik, enzimatik dan immunosurgery. Penelitian ini bertujuan mengamati efektifitas penggunanaan metode immunosurgery untuk memperoleh ICM dari embrio tahap blastosis. Embrio tahap blastosis diperoleh dari mencit betina strain Swiss Webster yang telah distimulasi menggunakan pregnant mare’s gonadotropin (PMSG) dan human chorionic gonadotropin (hCG). Inner Cell Mass (ICM) dikultur dan dilakukan pengamatan terhadap tingkat perlekatan ICM (attachment rate/AR) serta tingkat pembentukan koloni primer ESC (primary colony/PC) serta morfologi ESC line yang terbentuk. Hasil menunjukkan bahwa ICM yang diisolasi menggunakan metode immunosurgery memiliki AR dan PC masing-masing 78,57% dan 71,00%. Embryonic Stem Cell (ESC) dapat mengalami diferensiasi setelah beberapa pasase dengan membentuk embryoid body (EB). Penelitian ini menunjukkan bahwa metode immunosurgery merupakan metode yang efektif untuk memproduksi ESC line dari embrio mencit tahap blatosis.

Kata kunci : Blastosis, Inner cell mass, ICM, ESC