Skripsi

The Viability and Proliferation Capacity of Human Umbilical Cord Tissue Derived Mesenchymal Stem Cells in High Glucose Dulbecco’s Modified Eagle Medium (DMEM) Solution = The Viability and Proliferation Capacity of Human Umbilical Cord Tissue Derived Mesenchymal Stem Cells in High Glucose Dulbecco’s Modified Eagle Medium (DMEM) Solution.

Human umbilical-cord derived mesenchymal stem cells (hUC-MSCs) are commonly used in therapy for various diseases due to their multilineage differentiation and immunomodulation capacities. The current problem in usage is the delay in transplant often found due to administrative problems, following which the cells are sent back to the lab for re-culture, furthering the delay. This research aims to investigate whether High Glucose DMEM is a viable solution for temporary storage before transplant of cells. For this experiment, hUC-MSCs were suspended in High Glucose DMEM at 4 0 C. Outcomes of viability and proliferative capacity in terms of population doubling time were measured over 0, 3, 6, 24, 48, 72, 96, and 168 hours. Trypan Blue exclusion method was employed using a standard hemocytometer for viability and PDT calculations. Kruskal-Wallis test was employed to compare data to baseline. The study yielded that the viability of cells was maintained above 70% for up to 96 hours, but was significantly different as compared to baseline after 72 hours (P < 0.05). PDT was significantly different as compared to baseline after 24 hours (P < 0.05). In conclusion, High Glucose DMEM can be used as a temporary storage solution for hUC-MSCs for up to 24 hours after initial suspension. However, further experiments investigating the effects of suspension of cells in High Glucose DMEM on the differentiation and immunomodulation capacities of hUC-MSCs must be done.
Keywords: hUC-MSC, viability, PDT, High Glucose DMEM


Sel punca mesenkimal yang berasal dari tali pusat manusia (hUC-MSCs) umum digunakan dalam terapi berbagai penyakit karena pembelahan yang bersifat multipotensi dan kemampuan imunomodulasi. Masalah yang sekarang dihadapi dalam pelaksanaan transplantasi sel punca adalah keterlambatan karena proses administrasi. Akibatnya, sel punca tersebut harus dikembalikan ke laboratorium untuk dikultur ulang dan menyebabkan proses pelaksanaan transplantasi semakin terlambat. Studi ini bertujuan untuk mengetahui apakah larutan High Glucose DMEM dapat digunakan sebagai larutan penyimpanan sementara dari sel yang akan ditransplantasikan. Dalam percobaan ini, hUC-MSCs diletakkan dalam suspensi pada larutan High Glucose DMEM dalam suhu 4 o C. Hasil berupa viabilitas dan kapasitas proliferasi berdasarkan waktu penggandaan populasi (PDT) dari sel punca tersebut diukur pada jam ke-0, 3, 6, 24, 48, 72, 96, dan 168. Trypan Blue Exclusion Method dan alat hemositometer standar digunakan untuk mengukur viabilitas dan kalkulasi PDT. Uji Kruskal-Wallis digunakan untuk menganalisis data. Percobaan ini menghasilkan bahwa viabilitas dari sel punca dipertahankan hingga lebih dari 70% untuk 96 jam, namun berbeda makna setelah 72 jam (P < 0.05). PDT berbeda bermakna setelah 24 jam (P < 0.05). Untuk kesimpulan, larutan High Glucose DMEM dapat digunakan sebagai media penyimpanan sementara dari hUC-MSCs sampai selama 24 jam. Akan tetapi, dibutuhkan penelitian lanjutan untuk mengetahui kemampuan pembelahan dan imunomodulasi.
Kata kunci: hUC-MSC, viabilitas, PDT, High Glucose DMEM