Stem cells play a critical role in bone marrow transplant procedures because they are capable of differentiating into mature blood cells. New studies have shown that umbilical cord blood also contains large amounts of hematopoietic stem cells. As a result, both cord blood and bone marrow are now used for treating various immunological disorders, cancers, and specific genetic diseases.
This article demonstrates a technique for cord blood sample preparation and provides a precise set of instrument parameters for the Vi-CELL™ XR cell viability analyzer.
Bone Marrow Cells vs. Cord Blood Stem Cells
When compared to bone marrow cells, cord blood stem cells offer more benefits. For instance, stem cells isolated from cord blood can be easily accessed as they can be obtained from the placenta and umbilical cord during delivery. On the other hand, it is rather difficult to harvest stem cells from the bone marrow as it involves a surgical procedure.
For successful bone marrow transplantations, it is important to ensure that tissue proteins or antigens between the recipient and donor match closely. However, in case there is a slight mismatch, cord blood stem cells can still prove effective. As a result, recipients can significantly benefit from stem cells harvested from cord blood.
At present, there are a number of centers, which can be used by parents to store the cord blood of their newborn babies. When cells are stored in liquid nitrogen, two critical parameters must be properly assayed. These include percentage of cell viability, and cell concentration. These measurements are carried out before storage and after the thawing procedure. However, there is a possibility that both the number and viability of cells could reduce given that cryopreservative, usually dimethyl sulfoxide (DMSO), is used in the freezing process.
Beckman Coulter’s Vi-CELL™ XR (Figure 1) is specifically designed to automate the manual trypan blue, a vital dye exclusion technique used for determining cell viability. The instrument also allows unbiased measurements of cellular concentration.
Figure 1. The Vi-CELL XR
Methods and Materials
The SAS laboratory at Beckman Coulter obtained a cord blood sample from Miami-based Baptist Hospital. The blood was diluted in a ratio of 1:1 using phosphate buffered saline maintained at room temperature. The mononuclear cells were then separated using the Ficoll gradient separation method. The cells thus separated were cleaned in PBS and again suspended in 2mL of Isoflow (Isoton II).
Next, a cell suspension was prepared in a ratio of 1:10 by introducing 100µL of cells into 900µL of Isoflow in a Vi-CELL™ sample cup. The Vi-CELL™ XR was then employed to produce the cord blood cell type. The concentration control of the Vi-CELL™ XR was also assayed before sample analysis.
The results of the Vi-CELL concentration control are shown in Figure 2; a cord blood cell image on the Vi-CELL™ XR cell viability analyzer is shown in Figure 3; and the cell type parameters employed for cord blood cell analysis are illustrated in Figure 4. The size range was set between 3 and 25µm; imaging parameters are as follows:
- Cell sharpness: 100
- Cell brightness: 85
- Viable cell spot brightness: 65
- Viable cell spot area: 5
Figure 2. Results of the Vi-CELL concentration control
Figure 3. A cord blood cell image on the Vi-CELL™ XR
Figure 4. Cell type parameters used to analyze cord blood cells.
Infusions of cord blood stem cell offer tremendous benefits when compared to bone marrow transplants. Beckman Coulter’s Vi-CELL™ XR can be used for assaying both concentration and viability of cord blood cells. In this study, the cell type settings in case of cord blood samples were suitably determined.
About Beckman Coulter
Beckman Coulter develops, manufactures and markets products that simplify, automate and innovate complex biomedical tests. More than a quarter of a million Beckman Coulter instruments operate in laboratories around the world, supplying critical information for improving patient health and reducing the cost of care.
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