Tesis

Kadar bone morphogenetic protein-2, vascular endothelial growth factor, dan epidermal growth factor pada sekretom sel punca mesenkimal dari jaringan adiposa dan tali pusat = The level of bone morphogenetic protein-2, vascular endothelial growth factor, and epidermal growth factor in the secretome of adipose tissue and umbilical cordderived mesenchymal stem cells.

Pendahuluan: Pemberdayaan sekretom sel punca mesenkimal banyak diteliti belakangan ini karena dipercaya faktor pertumbuhan yang terkandung di dalamnya bekerja melalui mekanisme parakrin di situs cedera. Di antara banyaknya faktor-faktor pertumbuhan, beberapa disinyalir memiliki efek osteogenik antara lain bone morphogenetic protein-2 (BMP-2), epidermal growth factor (EGF), dan vascular endothelial growth factor (VEGF). Penelitian ini bertujuan untuk mengidentifikasi keberadaan BMP-2 yang belum pernah terdeteksi di dalam sekretom sel punca mesenkimal jaringan adiposa dan tali pusat, serta mengukur kadar EGF dan VEGF di dalamnya. Metode: Sampel sekretom dari sel punca mesenkimal jaringan adiposa dan tali pusat dibedakan berdasarkan perlakuan pemberian serum atau non serum dan waktu pengambilan saat penggantian medium terakhir atau saat panen, dan dianalisa dengan metode ELISA sandwich assay menggunakan Human BMP-2, VEGF, and EGF ELISA Kits. Hasil: Didapatkan 185 sampel sekretom jaringan adiposa dan 195 sampel sekretom jaringan tali pusat. Sebaran nilai BMP-2 tersentrasi pada nilai 0 pada sekretom jaringan adiposa maupun tali pusat. Kadar EGF dan VEGF memiliki perbedaan bermakna pada sampel jaringan adiposa yang berbeda (p < 0,009 dan p < 0,005). Kadar VEGF berkisar pada nilai 1473,5 pg/mL pada sekretom jaringan adiposa dan nilai 0 pg/mL pada sekretom jaringan tali pusat (p < 0,001). Pada sekretom jaringan adiposa, kadar EGF menurun seiring bertambahnya pasase (p < 0,009), sementara kadar VEGF meningkat seiring bertambahnya pasase (p < 0,001). Kesimpulan: Sekretom jaringan adiposa dan tali pusat kemungkinan hanya mengandung BMP-2 dalam nilai yang sangat rendah. Perbedaan donor sampel memberikan perbedaan kadar EGF dan VEGF pada sekretom jaringan adiposa yang diduga berkaitan dengan usia dan indeks massa tubuh donor. Kedua sekretom mengandung EGF namun hanya dalam jumlah moderat, dan hanya sedikit dipengaruhi oleh penambahan serum saat kultur. Kadar VEGF terdapat dalam jumlah tinggi dalam sekretom jaringan adiposa dan relatif konstan, dan sebaliknya hanya terkandung dalam jumlah yang sangat rendah pada sekretom jaringan tali pusat.
Kata kunci: Sekretom, media terkondisi, sel punca mesenkimal, faktor pertumbuhan


Introduction: The secretome derived from mesenchymal stem cells has recently garnered worldwide interests as it has been suggested that the growth factors contained within secretome works via a paracrine mechanism in the injured area. Of these factors, some are thought to have an osteogenic effect, including bone morphogenetic protein-2 (BMP-2), epidermal growth factor (EGF), and vascular endothelial growth factor (VEGF). Methods: Secretome samples from adipose tissue (AT-MSC) and umbilical cordderived mesenchymal stem cells (UC-MSC) were grouped based on serum administration and harvesting time, and were analyzed with an ELISA sandwich assay method using Human BMP-2, VEGF, and EGF ELISA Kits. This study aims to identify whether BMP-2 is contained in the secretome of AT-MSC and UC-MSC, which has never been reported before, and to measure the level of EGF and VEGF within the secretome. Results: There were 185 and 195 samples of secretome from AT-MSC and UCMSC, respectively. The distribution of value for BMP-2 was centered around zero in the secretome of both AT-MSC and UC-MSC. The level of EGF and VEGF were significantly different between different donor samples of AT-MSC (p < 0,009 and p < 0,005). The level of VEGF in the secretome of AT-MSC and UCMSC were approximately 1473,5 pg/mL and 0 pg/mL, respectively. On the ATMSC secretome, the level of EGF was found decreased with increasing passage (p < 0,009), whilst the level of VEGF was found increased with increasing passage (p < 0,001). Discussion: The secretome of AT-MSC and UC-MSC may contain BMP-2, despite being on a very low level. The difference in donor sample gave a significantly different level of EGF and VEGF on adipose tissue mesenchymal stem cells secretome, which might be related to the age and body mass index of the donor. Both adipose tissue and umbilical-cord derived secretome contained EGF but only in moderate amount, and was affected minimally by administration of serum. Level of VEGF was found high in adipose tissue mesenchymal stem cells secretome and was relatively constant, and conversely was contained on a very low level in umbilical cord-derived mesenchymal stem cells secretome.
Keywords: secretome, conditioned medium, mesenchymal stem cell, growth fact