The importance of amnioMAX medium in the recovery of human amniotic fluid cells from freezing at -80oC

Background: Recent developments in regenerative medicine led to a rapid rise in the use of stem cells and strategies for preservation and storage. Therefore, preparing a frozen stock of cells properly increases the validity of any results generated from the cell line.Objective: This paper examines the effect of medium on retrieving of human amniocytes from freezing at -80oC.Materials and Methods: Amniocytes were cultured in DMEM medium and then cells were frozen at passage 4 in FBS supplemented by 10% DMSO under the following conditions: 10 min at 4oC, 24 hr at -20oC and then cryopreserved in -80oC in four different time range including 4 months, 6 months, 8 months and 1 year, each group includes 3 samples. For recovery of the amniocytes, we used 3 protocols: AmnioMAX-II, DMEM and a modified medium composed of 2:1 v/v DMEM: AmnioMAX-II.Results: Our results have shown total recovery by three protocols in the group 4m. While in 6m group, only 2 samples in DMEM group could not be able to grow. However, the cells demonstrated incredibly lower viability in the time course of 8m and 1y, so that in the group DMEM only 1 sample in 8m and none of them in 1y category was able to proliferate. Nonetheless, this was amazingly different when we used AmnioMAX, as a results, recovery with AmnioMAX was 100% in all samples, from 4m to 1y of cryopreservation, even though DMEM:AmnioMAX-II (2:1 v/v) could recover only 2 samples in 8m and one sample in 1y.Conclusion: Our work has led us to conclude the importance of AmnioMAX medium in the recovery of human amniocytes in the long-term course of freezing at -80oC.

کلیدواژه ها:

Amniotic fluid cells ، Freezing procedure ، AmnioMAX ، Cryopreservation