Chyara Elisa Nofianti1, Intan Novita Sari1, Jeanne Adiwinata Pawitan2,3,4. 1. Faculty of Pharmacy, Andalas university, Padang, Indonesia. 2. Stem Cell Medical Technology Integrated Service Unit, Dr. Cipto Mangunkusumo General Hospital/Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia. 3. Department of Histology, Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia. 4. Stem Cell and Tissue Engineering Research Center, Indonesia Medical Education and Research Institute (IMERI), Faculty of Medicine Universitas Indonesia, Jakarta, Indonesia.
Abstract
BACKGROUND: We have developed a simple lipoaspirate washing method using a coffee filter to eliminate liposuction noxious material before isolating adipose tissue derived mesenchymal stem cells (AT-MSCs), and used them in clinical trials. Before administration to patients, MSCs are usually suspended in physiologic saline. However, MSCs only survive for a limited time in physiologic saline. Therefore, alternative solution that can preserve MSC survival will be beneficial. Therefore, the purpose of this study was to compare the use of physiologic saline and Dulbecco's modified Eagle's Medium (DMEM) as temporary storage solution for our AT-MSCs. METHODS: We did viability assessments of AT MSCs after 0, 3, 6, 24, 48, 72, and 96 hours suspended in physiologic saline compared to DMEM, and stored at 4 ℃. Further proliferation capacities of the cells after various suspension times were assessed. All viability and proliferation capacity assessments were done in four replications. Differences between the various suspension time in terms of viability and proliferation capacity were compared and tested by ANOVA or Kruskal-Wallis test. RESULTS: Viability was >70% after 48 hours in physiologic saline and 24 hours in DMEM, which showed that physiologic saline was superior compared to DMEM. Increase in PDT began to be significant compared to initial PDT after 24 hours in both physiologic saline, and DMEM. CONCLUSIONS: For our AT-MSCs, physiologic saline was superior to DMEM, and storage should not exceed 24 hours.
BACKGROUND: We have developed a simple lipoaspirate washing method using a coffee filter to eliminate liposuction noxious material before isolating adipose tissue derived mesenchymal stem cells (AT-MSCs), and used them in clinical trials. Before administration to patients, MSCs are usually suspended in physiologic saline. However, MSCs only survive for a limited time in physiologic saline. Therefore, alternative solution that can preserve MSC survival will be beneficial. Therefore, the purpose of this study was to compare the use of physiologic saline and Dulbecco's modified Eagle's Medium (DMEM) as temporary storage solution for our AT-MSCs. METHODS: We did viability assessments of AT MSCs after 0, 3, 6, 24, 48, 72, and 96 hours suspended in physiologic saline compared to DMEM, and stored at 4 ℃. Further proliferation capacities of the cells after various suspension times were assessed. All viability and proliferation capacity assessments were done in four replications. Differences between the various suspension time in terms of viability and proliferation capacity were compared and tested by ANOVA or Kruskal-Wallis test. RESULTS: Viability was >70% after 48 hours in physiologic saline and 24 hours in DMEM, which showed that physiologic saline was superior compared to DMEM. Increase in PDT began to be significant compared to initial PDT after 24 hours in both physiologic saline, and DMEM. CONCLUSIONS: For our AT-MSCs, physiologic saline was superior to DMEM, and storage should not exceed 24 hours.
Entities:
Keywords:
Adipose tissue; mesenchymal stem cell (MSC); population doubling time (PDT); storage solution; viability
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