Patients worldwide require therapeutic transfusions of packed red blood cells (pRBCs), which is applied to the high-risk patients who need periodic transfusions due to leukemia, lymphoma, myeloma and other blood diseases or disorders. Contrary to the general hospital population where the transfusions are carried out mainly for healthy trauma patients, in case of high-risk patients the proper quality of pRBCs is crucial. This leads to an increased demand for efficient technology providing information on the pRBCs alterations deteriorating their quality. Here we present the design of an innovative, label-free, noninvasive, rapid Raman spectroscopy-based method for pRBCs quality evaluation, starting with the description of sample measurement and data analysis, through correlation of spectroscopic results with reference techniques' outcomes, and finishing with methodology verification and its application in clinical conditions. We have shown that Raman spectra collected from the pRBCs supernatant mixture with a proper chemometric analysis conducted for a minimum one ratio of integral intensities of the chosen Raman marker bands within the spectrum allow evaluation of the pRBC quality in a rapid, noninvasive, and free-label manner, without unsealing the pRBCs bag. Subsequently, spectroscopic data were compared with predefined reference values, either from pRBCs expiration or those defining the pRBCs quality, allowing to assess their utility for transfusion to patients with acute myeloid leukemia (AML) and lymphoblastic leukemia (ALL).
In conclusion, we have shown that Raman spectra collected for SM with a proper chemometric analysis conducted for minimum one ratio of integral intensities of the chosen Raman marker bands allow to define the pRBCs quality in a rapid, non-invasive, free-label manner, without opening the pRBCs bag. The results of the analysis are compared to the predefined spectroscopic reference values, either from pRBCs expiration or ones specific to the pRBCs quality, to assess its utility for transfusion to AML and ALL patients. Moreover, there are various conditions such as hemophilia, thalassemia, sickle diseases, etc., that requires blood transfusion where the method presented herein would be useful to assess pRBCs quality rapidly prior to the transfusion.
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To perform ex vivo studies, human cord blood units were provided by the CORD:USE Cord Blood Bank (Orlando, FL, USA). All cord blood samples were screened for alanine aminotransferase (ALT) and pathogenic antigen antibodies (including anti-HCV, anti-HBsAg, anti-HIV, anti-Syphilis, anti-Chlamydia, and anti-Gonorrhea Abs), and only pathogen-free cord blood units were used for isolating CB-SCs. Human cord blood-derived stem cells (CB-SCs) were generated as previously described [24,25] with the following modifications. Cord blood mononuclear cells were plated in serum-free culture medium (Lonza, Walkersville, MD, USA) and incubated at 37C, in 8% CO2. After 2 to 3 weeks, CB-SCs growing at 80-90% confluence were prepared for co-culture with allogeneic lymphocytes. 2ff7e9595c
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