PERAN HIPOKSIA TERHADAP PERTUMBUHAN DAN DIFERENSIASI HUMAN ADIPOSE DERIVED STEM CELLS YANG DITUMBUHKAN PADA SCAFFOLD SUTRA DIINDUKSI OLEH PLATELET RICH PLASMA

• Main Author: Widya Sarjana, Ergha
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/43462
• Institution: Institut Teknologi Bandung

Stem cells in tissue engineering have a high potential to cure several diseases such as low back pain (LBP). LBP is commonly caused by the degradation of a part in the intervertebral disc (IVD) called nucleus pulposus (NP). Inside the human body, NP is found in hypoxic condition and containing a lot of chondrocytes with extracellular matrices such as type II collagen and glycosaminoglycans (GAG). Tissue engineering of human adipose derived stem cells (hADSC), which come from fat tissue, has a high potential to be used for LBP therapy because of its ability to be differentiated as chondrocyte. Previous research has proven that hADSC grown in silk scaffold with 500?m pores and 10% induction of platelet-rich plasma (PRP) could increase the growth and differentiation of hADSC to become chondrocyte. In accordance to the body’s conditions, low oxygen concentration is an essential factor that can transform hADSC ability to grow and differentiate due to its ability to activate a transcription factor called the hypoxiainducible factor-1? (HIF-1?). This research is conducted to analyze the difference in hADSC ability to grow within normoxic and hypoxic conditions, also the difference in hADSC ability to differentiate within normoxic and hypoxic conditions observed from the number of GAG presence and the visualization of type II collagen and HIF-1?. The stem cells coming from fat tissue is initially characterized by plastic adherence test, hADSC marker presence, and multipotency test. HADSC ability to grow is determined by MTT Assay. HADSC differentiation is tested with analysis of the GAG quantity using alcian blue coloring method and immunocytochemistry (ICC) method. The cells used in this research, which obtained from the fat tissue, has met the requirements from the three of hADSC characterization test so it is surely confirmed as hADSC. The hADSC is cultured on a silk scaffold with 500 ?m pores and 10% PRP induction in normoxic, 2% hypoxic and 5% hypoxic conditions for 21 days. Higher type II collagen visualization was detected in hypoxic conditions than in normoxic condition. HIF-1? is only detected in hADSC cultured in hypoxic conditions. hADSC ability to grow is higher in hypoxic conditions than in normoxic condition, but the cell grown in hypoxic condition declines after 7 days. hADSC ability to differentiate is higher in hypoxic conditions than in normoxic condition based on the number of GAG observed. This research concludes that hypoxia could increase hADSC ability to grow and differentiated as chondrocyte.