Quantification of Antibody Persistence for Cell Surface Protein Labeling

Dempsey, Megan E.; Woodford-Berry, Olivia; Darling, Eric M.

doi:10.1007/s12195-021-00670-3

Cited by 4 publications

(2 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In addition, the use of antibodies can cause changes in the structure of the cell membrane, and subsequently an additional step of detachment and recovery of the used antibodies is required. 10 The general protocol used for ASC extraction is shown in Fig. 2.…”

Section: Extraction Methodsmentioning

confidence: 99%

“…However, the high investment and operating costs, as a consequence of the equipment and reagents they require, have limited their use at a laboratory scale. In addition, the use of antibodies can cause changes in the structure of the cell membrane, and subsequently an additional step of detachment and recovery of the used antibodies is required 10 …”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Aqueous two‐phase systems as a novel alternative in the primary recovery of human mesenchymal cells obtained from adipose tissue

Freire‐Flores,

Llanovarced‐Kawles,

Andrews

et al. 2024

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

BACKGROUND: In recent years, therapies based on regenerative medicine have gained wide interest due to their potential in organ transplantation and in the treatment of multiple diseases. These cell‐based therapies rely mainly on the therapeutic potential of stem cells due to their self‐renewal, differentiation, capacities and immunosuppressive properties. Adipose tissue contains a large number of mesenchymal stem cells compared to other tissues and is easily accessible through non‐invasive techniques; hence the primary recovery of adipose‐derived mesenchymal stem cells (ASC) becomes a critical step. Aqueous two‐phase systems (ATPS) have shown promise as a separation technique for mesenchymal stem cells (MSC) due to their scalability, cost‐effectiveness, and compatibility. RESULTS: In this study, we investigated the partitioning behavior of the stromal vascular fraction cells, including ASC, in different ATPS compositions (polyethylene glycol, dextran, ficoll and Ucon). Among the three ATPS tested, most of the ASC consistently migrate to the top phase as indicated by immunostaining studies for CD73, CD90 and CD105 stem cell markers. Nevertheless, the Ficoll‐DEX system retrieves the greatest quantity of ASC recovered in the top phase (89% of total ASC), while the Ucon‐DEX system effectively eliminates the most contaminating cells (38%). CONCLUSIONS: Our findings provide insights into the conditions under which ASC and contaminant cells concentrate in opposite phases in different ATPS, which can contribute to the development of efficient and scalable separation systems for MSC in regenerative medicine and tissue engineering applications.This article is protected by copyright. All rights reserved.

show abstract

Section: Extraction Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Aqueous two‐phase systems as a novel alternative in the primary recovery of human mesenchymal cells obtained from adipose tissue

Freire‐Flores,

Llanovarced‐Kawles,

Andrews

et al. 2024

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

show abstract

Discovery of surface biomarkers for cell mechanophenotype via an intracellular protein-based enrichment strategy

Dempsey

Chickering

González-Cruz

et al. 2022

Cell. Mol. Life Sci.

View full text Add to dashboard Cite

Optimized Enrichment of Murine Blood-Brain Barrier Vessels with a Critical Focus on Network Hierarchy in Post-Collection Analysis

Abdelazim,

Barnes,

Stupin

et al. 2024

Preprint

View full text Add to dashboard Cite

Cerebrovascular networks contain a unique region of interconnected capillaries known as the blood-brain barrier (BBB). Positioned between upstream arteries and downstream veins, these microvessels have unique structural features, such as the absence of vascular smooth muscle cells (vSMCs) and a relatively thin basement membrane, to facilitate highly efficient yet selective exchange between the circulation and the brain interstitium. This vital role in neurological health and function has garnered significant attention from the scientific community and inspired methodology for enriching BBB capillaries. Extensive characterization of the isolates from such protocols is essential for framing the results of follow-on experiments and analyses, providing the most accurate interpretation and assignment of BBB properties. Seeking to aid in these efforts, here we visually screened output samples using fluorescent labels and found considerable reduction of non-vascular cells following density gradient centrifugation (DGC) and subsequent filtration. Comparatively, this protocol enriched brain capillaries, though larger diameter vessels associated with vSMCs could not be fully excluded. Protein analysis further underscored the enrichment of vascular markers following DGC, with filtration preserving BBB-associated markers and reducing, though not fully removing, arterial/venous contributions. Transcriptional profiling followed similar trends of DGC plus filtration generating isolates with less non-vascular and non-capillary material included. Considering vascular network hierarchy inspired a more comprehensive assessment of the material yielded from brain microvasculature isolation protocols. This approach is important for providing an accurate representation of the cerebrovascular segments being used for data collection and assigning BBB properties specifically to capillaries relative to other regions of the brain vasculature.

show abstract

Quantification of Antibody Persistence for Cell Surface Protein Labeling

Cited by 4 publications

References 43 publications

Aqueous two‐phase systems as a novel alternative in the primary recovery of human mesenchymal cells obtained from adipose tissue

Aqueous two‐phase systems as a novel alternative in the primary recovery of human mesenchymal cells obtained from adipose tissue

Discovery of surface biomarkers for cell mechanophenotype via an intracellular protein-based enrichment strategy

Optimized Enrichment of Murine Blood-Brain Barrier Vessels with a Critical Focus on Network Hierarchy in Post-Collection Analysis

Contact Info

Product

Resources

About