Improved cord blood thawing procedure enhances the reproducibility and correlation between flow cytometry CD34 +  cell viability and clonogenicity assays

Galindo, Cristian Camilo; Lozano, Diana María Vanegas; Rodríguez, Bernardo Camacho; Perdomo-Arciniegas, Ana-María

doi:10.1016/j.jcyt.2018.03.033

Cited by 5 publications

(4 citation statements)

References 8 publications

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“…UCB units bear three attached segments (distal, medial, and proximal according to their proximity to the unit), which are considered representative samples to verify cell viability, cell recovery, and clonogenic potency (CLONE) after thawing. This process is denominated confirmatory testing and the protocol was previously standardized in the bank for clinical purposes [27]. Confirmation of the distal segment as a representative sample of the UCB unit was also performed to guarantee reliability of the results [28].…”

Section: Resultsmentioning

confidence: 99%

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Human Leukocyte Antigen and Red Blood Cells Impact Umbilical Cord Blood CD34+ Cell Viability after Thawing

Vanegas¹,

Galindo²,

Páez-Gutiérrez³

et al. 2019

IJMS

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Hematopoietic progenitor cell (HPC) transplantation is a treatment option for malignant and nonmalignant diseases. Umbilical cord blood (UCB) is an important HPC source, mainly for pediatric patients. It has been demonstrated that human leukocyte antigen (HLA) matching and cell dose are the most important features impacting clinical outcomes. However, UCB matching is performed using low resolution HLA typing and it has been demonstrated that the unnoticed mismatches negatively impact the transplant. Since we found differences in CD34+ viability after thawing of UCB units matched for two different patients (p = 0.05), we presumed a possible association between CD34+ cell viability and HLA. We performed a multivariate linear model (n = 67), comprising pre-cryopreservation variables and high resolution HLA genotypes separately. We found that pre-cryopreservation red blood cells (RBC), granulocytes, and viable CD34+ cell count significantly impacted CD34+ viability after thawing, along with HLA-B or -C (R2 = 0.95, p = 0.01; R2 = 0.56, p = 0.007, respectively). Although HLA-B*40:02 may have a negative impact on CD34+ cell viability, RBC depletion significantly improves it.

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Section: Resultsmentioning

confidence: 99%

“…All samples were thawed using the previously mentioned standardized protocol [27]. CD34 + cell viability after thawing exhibited a statistically significant difference between UCB unit distal segment and RBC-depleted MNC ( p < 0.0001).…”

Section: Resultsmentioning

confidence: 99%

Human Leukocyte Antigen and Red Blood Cells Impact Umbilical Cord Blood CD34+ Cell Viability after Thawing

Vanegas¹,

Galindo²,

Páez-Gutiérrez³

et al. 2019

IJMS

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“…The CVS assay principle is based on measuring sample adherence by coloring attached samples with CVS, a protein and DNA binding dye [ 52 ]. The principle behind the colonogenic assay, also known as the plating assay, is that live samples will generate colonies that are easy to observe for which the number of surviving samples can be easily estimated [ 53 , 54 , 55 ]. LDH is released into the extracellular space when the plasma membrane is disrupted, which can be a significant indicator of necrotic cells [ 56 , 57 ].…”

Section: Living Sample Viability Measurement Methodsmentioning

confidence: 99%

Living Sample Viability Measurement Methods from Traditional Assays to Nanomotion

Al-Madani

Yao

et al. 2022

Biosensors

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Living sample viability measurement is an extremely common process in medical, pharmaceutical, and biological fields, especially drug pharmacology and toxicology detection. Nowadays, there are a number of chemical, optical, and mechanical methods that have been developed in response to the growing demand for simple, rapid, accurate, and reliable real-time living sample viability assessment. In parallel, the development trend of viability measurement methods (VMMs) has increasingly shifted from traditional assays towards the innovative atomic force microscope (AFM) oscillating sensor method (referred to as nanomotion), which takes advantage of the adhesion of living samples to an oscillating surface. Herein, we provide a comprehensive review of the common VMMs, laying emphasis on their benefits and drawbacks, as well as evaluating the potential utility of VMMs. In addition, we discuss the nanomotion technique, focusing on its applications, sample attachment protocols, and result display methods. Furthermore, the challenges and future perspectives on nanomotion are commented on, mainly emphasizing scientific restrictions and development orientations.

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“…However, the cells expanded ex vivo undergo changes in genetic expression, which is an issue that requires further studies [119]. Additionally, the parameters of CB units may be ameliorated by improving the CB thawing procedure [120], using DMSO dextrose instead of DMSO [121], and adding new substances before cryopreservation, such as quercetin [122].…”

Section: Past and Future Perspectivesmentioning

confidence: 99%

Autologous Cord Blood in Children with Cerebral Palsy: A Review

Boruczkowski

Pujal²,

Zdolińska-Malinowska

2019

IJMS

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The aim of this narrative review is to report on the current knowledge regarding the clinical use of umbilical cord blood (CB) based on articles from PubMed and clinical trials registered on ClinicalTrials.gov. An increasing amount of evidence suggests that CB may be used for both early diagnostics and treatment of cerebral palsy. The acidity of CB and its biochemical parameters, including dozens of cytokines, growth factors, and other metabolites (such as amino acids, acylcarnitines, phosphatidylcholines, succinate, glycerol, 3-hydroxybutyrate, and O-phosphocholine) are predictors of future neurodevelopment. In addition, several clinical studies confirmed the safety and efficacy of CB administration in both autologous and allogeneic models, including a meta-analysis of five clinical trials involving a total of 328 participants. Currently, nine clinical trials assessing the use of autologous umbilical CB in children diagnosed with hypoxic-ischemic encephalopathy or cerebral palsy are in progress. The total population assessed in these trials exceeds 2500 patients.

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Improved cord blood thawing procedure enhances the reproducibility and correlation between flow cytometry CD34 + cell viability and clonogenicity assays

Cited by 5 publications

References 8 publications

Human Leukocyte Antigen and Red Blood Cells Impact Umbilical Cord Blood CD34+ Cell Viability after Thawing

Human Leukocyte Antigen and Red Blood Cells Impact Umbilical Cord Blood CD34+ Cell Viability after Thawing

Living Sample Viability Measurement Methods from Traditional Assays to Nanomotion

Autologous Cord Blood in Children with Cerebral Palsy: A Review

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