Validation of the single-platform ISHAGE protocol for enumeration of CD34+ hematopoietic stem cells in umbilical cord blood in a Brazilian center

New drug targets, markers of disease prognosis, and more efficient treatment options are an unmet clinical need in breast cancer (BC). We have conducted a pilot study including patients with luminal B stage breast cancer IIA–IIIB. The presence and frequency of various populations of cancer stem cells (CSC) and somatic stem cells were assessed in the blood, breast tumor tissue, and normal breast tissue. Our results suggest that patients with BC can be divided into two distinct groups based on the frequency of aldehyde dehydrogenase positive cells (ALDH1+ cells) in the blood (ALDH1hi and ALDH1low). In the ALDH1hi cells group, the tumor is dominated by epithelial tumor cells CD44+CD24low, CD326+CD44+CD24−, and CD326−CD49f+, while in the ALDH1low cells group, CSCs of mesenchymal origin and epithelial tumor cells (CD227+CD44+CD24− and CD44+CD24−CD49f+) are predominant. In vitro CSCs of the ALDH1low cells group expressing CD326 showed high resistance to cytostatics, CD227+ CSCs of the ALDH1hi cells group are sensitive to cytostatics. Epithelial precursors of a healthy mammary gland were revealed in normal breast tissue of patients with BC from both groups. The cells were associated with a positive effect of chemotherapy and remission in BC patients. Thus, dynamic control of their presence in blood and assessment of the sensitivity of CSCs to cytostatics in vitro can improve the effectiveness of chemotherapy in BC.

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“…The reagent contains FITC-labeled CD45, clone 2D1, and PE-labeled CD34, clone 8G12. We determined HSC according to the ISHAGE protocol [18].…”

Section: Detection Of Hemopoietic Stem Cellsmentioning

confidence: 99%

Cancer Stem Cells and Somatic Stem Cells as Potential New Drug Targets, Prognosis Markers, and Therapy Efficacy Predictors in Breast Cancer Treatment

et al. 2021

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“… 1 – 3 CD34+ cell content is a widely used predictive factor for accessing engraftment and survival following umbilical cord blood transplantation. 4 – 6 CD34 is expressed on HSCs and is considered a reliable marker for stem cells with regenerative capacity. 7 Despite the major limitation of lower HSCs, 8 with a single cord blood unit containing approximately 10% fewer stem cells than bone marrow, the number of CD34+ cells obtained per unit volume is similar.…”

Section: Introductionmentioning

confidence: 99%

Enumeration of viable CD34+ cells in cord blood using a novel stem cell enumeration kit

Huang

et al. 2021

J Int Med Res

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Objective To assess the detection performance of the hematopoietic stem cell enumeration kit developed by BD Biosciences. Methods Cord blood samples were prepared using a hematopoietic stem cell enumeration kit developed by BD Biosciences and Stem-Kit reagents from Beckman Coulter. CD34+ cells were enumerated using a BD FACSCanto instrument and FACSDiva software. Results A total of 519 samples were analyzed in this study. The hematopoietic stem cell enumeration kit developed by BD Biosciences yielded absolute counts of CD34-positive cells that were on average 8.7% lower than Beckman Coulter Stem-Kit reagents (range: −5.7% to−14.7%). The BD Biosciences kit yielded relative counts that were on average 9.9% higher compared with Beckman Coulter Stem-Kit reagents (range: −2.1% to +13.8%). The intraclass correlation coefficients for absolute and relative counts of CD34-positive cells were 0.9967 (95% confidence interval [CI]: 0.9961–0.9972) and 0.9512 (95% CI: 0.9423–0.9587) for the BD Biosciences and Beckman Coulter kits, respectively. Conclusions The hematopoietic stem cell enumeration kit developed by BD Biosciences can be used to enumerate CD34-positive stem cells from cord blood samples.

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The single-cell big data analytics

Rawat,

Sharma,

Mohanty

2024

Computational Biology for Stem Cell Research

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Validation of the single-platform ISHAGE protocol for enumeration of CD34+ hematopoietic stem cells in umbilical cord blood in a Brazilian center

Cited by 4 publications

References 19 publications

Cancer Stem Cells and Somatic Stem Cells as Potential New Drug Targets, Prognosis Markers, and Therapy Efficacy Predictors in Breast Cancer Treatment

Cancer Stem Cells and Somatic Stem Cells as Potential New Drug Targets, Prognosis Markers, and Therapy Efficacy Predictors in Breast Cancer Treatment

Enumeration of viable CD34+ cells in cord blood using a novel stem cell enumeration kit

The single-cell big data analytics

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