Effect of Platelet Rich Plasma on Post Cryopreservation Viability, Morphology and Proliferation of Human Umbilical Cord Stem Cells

Abstract

In most cryopreservation medium, Fetal Bovine Serum (FBS) is used as supplement, while it is well known that FBS contains xenoproteins that can be incorporated into the cells and may be harmfull, as they can elicit immune response. Therefore, finding other xenofree materials as FBS alternative in cryopreservation medium is very important. Platelet Rich Plasma (PRP) is albumin rich and is a candidate for FBS alternative as cryopreservation supplement. Albumin is a natural extracellular cryoprotective agent that stabilizes impaired cell membrane during cryopreservation. This was an in vitro analytical study to to compare the effect of PRP and FBS as supplement in cryopreservation medium on human umbilical cord stem cells. In this study the stem cells were isolated from an umbilical cord tissue by explant method and propagated untill we got enough cells for cryopreservation. Cryopreservations were done using eight types of protocol, which differed in type and concentration of supplement and cell concentration. The effect of the eight protocols were compared in terms of post cryopreservation cell viability, morphology, cell size and proliferation. There were no difference between FBS and PRP supplemented cryopreservation media in term of cell viability and morphology. PRP supplemented medium showed better post cryopreservation performance in cell size and proliferation. PRP can be used as an alternative to FBS in cryopreservation medium for human umbilical cord tissue derived stem cells.

Effect of Cryopreservation and cumulative population doublings on Senescence of Umbilical Cord Mesenchymal Stem Cells

“…9 Our previous result showed that serial passages after cryopreservation in the same method as this research caused enlargement of cell size at P-2 compared to fresh cells (mean value 2674 vs 2056 µm 2 ), 11 though this study showed that in P-2 there was no senescent cells. Therefore, after cryopreservation, P-2 cells in this study might begin the senescent process, but had not accumulated enough β galactosidase to be detected with β galactosidase staining, which in our study, began to detect senescence in P-3.…”

“…Cell density at cryopreservation was 500,000 cells/mL, and cryopreservation procedure was done at -20 o C for 24 hours, and then moved into liquid N2 tank, vapor phase as described previously. 11 After one month, the cells were thawed and subjected to serial passages until P-8, and senescent analysis was done for all passages.…”

“…11 In brief, the cryotubes were immersed in a 37 o C water bath. After that, the cells were transferred into a complete medium, washed, and viable and total cells were counted by trypan blue exclusion method.…”

Comparison of in Vitro Vasculogenesis Potential between ASCs with BM-MSCs

“…This research was conducted for 16 months, from August 2016 -October 2017. The samples of this study were mesenchymal stem cells from bone marrow isolated by the simple method by Pawitan et al [36] and mesenchymal stem cells from fat tissue isolated by the simple method belonging to the RSCM-FKUI UPT Stem Cell Laboratory [37] . All cells were cryopreserved in the first phase using the xeno-free cryopreservation method.…”