Abstract
BackgroundThe finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation. Due to the once-in-a-life time chance of collecting it, an optimum cryopreservation method that can preserve the life and function of the cells contained is critically needed.MethodsUntil now, slow-cooling has been the routine method of cryopreservation; however, rapid-cooling offers a simple, efficient, and harmless method for preserving the life and function of the desired cells. Therefore, this study was conducted to compare the effectiveness of slow- and rapid-cooling to preserve umbilical cord blood of mononucleated cells suspected of containing hematopoietic stem cells. The parameters used in this study were differences in cell viability, malondialdehyde content, and apoptosis level. The identification of hematopoietic stem cells themselves was carried out by enumerating CD34+ in a flow cytometer.ResultsOur results showed that mononucleated cell viability after rapid-cooling (91.9%) was significantly higher than that after slow-cooling (75.5%), with a p value = 0.003. Interestingly, the malondialdehyde level in the mononucleated cell population after rapid-cooling (56.45 μM) was also significantly higher than that after slow-cooling (33.25 μM), with a p value < 0.001. The apoptosis level in rapid-cooling population (5.18%) was not significantly different from that of the mononucleated cell population that underwent slow-cooling (3.81%), with a p value = 0.138. However, CD34+ enumeration was much higher in the population that underwent slow-cooling (23.32 cell/μl) than in the one that underwent rapid-cooling (2.47 cell/μl), with a p value = 0.001.ConclusionsRapid-cooling is a potential cryopreservation method to be used to preserve the umbilical cord blood of mononucleated cells, although further optimization of the number of CD34+ cells after rapid-cooling is critically needed.
Highlights
The finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation
Our results were similar to those stated in research by Boyse et al, which showed an overall value of cell viability after cryopreservation with the slow-cooling method ranging from 13.5% to 67.6% [26]
Other results reported by Bayer et al clearly demonstrated that the viability of hematopoietic stem cells after cryopreservation using the conventional slow-cooling method was very low, with a mean value of 62% and an overall value ranging from 51% to 64% [27]
Summary
The finding of human umbilical cord blood as one of the most likely sources of hematopoietic stem cells offers a less invasive alternative for the need of hematopoietic stem cell transplantation. Further research about the properties of human umbilical cord blood led to reports that there are some types of stem cells in umbilical cord blood mononucleated cells, including hematopoietic stem cells. At least three sources are known for their potential to become the source of hematopoietic stem cells, i.e., the bone marrow, umbilical cord, and peripheral blood. Bone marrow is the most commonly used source of hematopoietic stem cells for transplantation. It has been proven to be quite effective, the collection of hematopoietic stem cells from bone marrow is an invasive and traumatic procedure. The use of peripheral blood requires a preliminary procedure, such as the administration of the granulocyte colony stimulating factor (G-CSF) prior to collection
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