Skripsi

The expression of Sox2 gene in adipose-derived stem cells and umbilical cord-derived stem cells, a comparison study = Ekspresi gen Sox2 pada Adipose Derived Stem Cell (ADSC) dan Umbilical Cord Stem Cell (USC), sebuah studi perbandingan.

Background: Ever since scientists successfully developed a method to derive stem cells from human embryos in 1998, stem cell research has been gaining momentum and nothing seems to indicate that this field of science will slow down anytime soon. To date, stem cells have been identified from 2 different sources: embryonic stem cells and from somatic tissues (somatic stem cells) – these two sources generate stem cells that differ in their potency. Embryonic stem cells (ESC), are pluripotent, making them the ideal stem cell that could be used in stem cell therapies. Its usage in such therapies are still rarely found, however, because of ethical reasons. Therefore, the possibilities of using other sources of stem cells, including Adipose Derived Stem Cells and Umbilical Cord Derived Stem Cells are currently being studied. Since the knowledge regarding the pluripotency of these two cells are still limited, this research aims to analyze the pluripotency level of the two type of stem cells, compared through the expression of Sox2, a transcriptional factor important in the maintenance of pluripotency. Method: RNA was extracted from USC and ADSC samples. One-Step Real-Time RT-PCR was then performed in order to calculate the relative expression of Sox2 gene in both samples to determine the pluripotency level of each cell. Electrophoresis was done to confirm the amplified sequence in One-Step RealTime RT-PCR. Results: Sox2 is found to be expressed only 1.10 times more highly in USC than in ADSC, which is not enough to indicate significant results. Conclusion: USC and ADSC have similar levels of Sox2 relative gene expression. There is no significant difference found between the two, thus, these two samples have similar pluripotency, making them both attractive sources of stem cells.
Keywords: adipose derived stem cell, umbilical cord stem cell, Sox2, gene expression


Latar Belakang: Sejak para ilmuwan telah sukses dalam mengekstraksi sel punca dari embrio manusia pada tahun 1998, riset mengenai sel punca menjadi sangat popular hingga sekarang. Kini, sel punca dapat dibedakan menjadi dua: Embryonic Stem Cells (ESC) dan Somatic Stem Cells – dua tipe sel punca ini berbeda dalam kepuncaan mereka. Embryonic Stem Cell (ESC) adalah sel punca yang pluripoten, hal ini membuat ESC menjadi kandidat yang ideal untuk terapi regeneratif menggunakan sel punca. Akan tetapi, ESC jarang digunakan karena konsiderasi etis. Maka dari itu, kemungkinan menggunakan sel punca lainnya, termasuk Adipose Derived Stem Cells (ADSC) dan Umbilical Cord Derived Stem Cells (USC) sedang dipelajari. Sebab informasi mengenai pluripotency dari kedua sel punca ini masih terbatas, penelitian ini bertujuan untuk menganalisa tingkat pluripotency dari ADSC dan USC, melalui ekspresi gen Sox2, salah satu faktor transkripsi yang penting dalam menjaga pluripotency suatu sel. Metode: RNA diekstraksi dari ADSC dan USC. One-Step Real-Time RT-PCR dilakukan untuk mendapatkan ekspresi relatif gen Sox2 yang menggambarkan tingkat pluripotency sel induk tersebut. Kemudian, electrophoresis dilakukan untuk mengkonfirmasi hasil amplifikasi gen dari One-Step Real-Time RT-PCR. Hasil: Ekspresi relatif gen Sox2 ditemukan 1.10 kali lebih tinggi pada USC dibandingkan ADSC, suatu perbedaan yang tidak cukup signifikan. Konklusi: USC dan ADSC memiliki tingkat ekspresi gen Sox2 relatif yang serupa. Tidak ada perbedaan yang signifikan yang ditemukan diantara keduanya, sehingga dapat disimpulkan bahwa kedua sel ini memiliki tingkat pluripotency yang mirip, membuat keduanya sumber sel punca yang sesuai.
Kata kunci: adipose derived stem cell, umbilical cord stem cell, Sox2, gene expression