Selection of DNA Aptamers for Differentiation of Human Adipose-Derived Mesenchymal Stem Cells from Fibroblasts

Abstract

In recent years, stem cell therapy has shown promise in regenerative medicine. In this context, the distinction of mesenchymal stem cells from broblasts has been crucial for safe clinical application of these cells. In the present study, we developed aptamers capable of speci cally recognize mesenchymal stem cells derived from adipose tissue (ASC) using the Cell-SELEX technique. We tested the a nity of ASC aptamers compared to dermal broblasts (FIB).Quantitative PCR was advantageous for the validation of four candidate aptamers in vitro. The binding capabilities of Apta 2 and Apta 42 could not distinguish both cell types, while Apta 21 and Apta 99 showed a better binding capacity to ASC with dissociation constants (Kd) of 50.46 ± 2.28 nM and 72.71 ± 10.3 nM, respectively. However, Apta 21 showed a Kd of 86.78 ± 9.14 nM when incubated with FIB. Therefore, only Apta 99 showed speci city to detect ASC. This aptamer is a promising tool for the in vitro identi cation of ASC. These results will help understand the differences between these two cell types for more speci c and precise cell therapies.

Adipose-Derived Stromal Cells for Chronic Wounds: Scientific Evidence and Roadmap Toward Clinical Practice

“…Although not considered crucial, the multipotent capacities of ASC could play a role in their therapeutic action on wounds. ASC can indeed differentiate into dermic fibroblasts accelerating granulation tissue formation 30 , 31 or keratinocyte-like cells having the capacity to form a stratified epidermis. 32 ASC are enriched in the stromal vascular fraction (SVF) of the adipose tissue, which can be directly used for therapeutic intervention.…”

Synthesis and characterization of gold nanorods coated by mesoporous silica MCM-41 as a platform for bioapplication in photohyperthermia

“…(3) there is less out of focus fluorescence collected when compared to a confocal microscope [93,94]. Although more extensive and conclusive biological assays must be performed in the future, these results suggest that the nanocomposites can be adhered to the cell membranes or internalized by the cells.…”

“…Depending on the excitation wavelength, the objective numerical aperture (N.A.) and the thickness of the excitation depth, which is defined as the evanescent field, can be less than 100 nm from the coverslip [92,93]. The advantages of such a restricted volume of illumination are as follows: (1) less photobleach; (2) the background is less than 2000 times when compared to standard epifluorescence microscopes; and…”

Application of aptamers in regenerative medicine

“…They have shown the ability to capture and enrich corresponding stem cells in bone marrow, whole blood, and other cell suspensions, suggesting their potential application in the collection of exogenous stem cells before transplantation and surface modification of biomaterials for tissue regeneration. Recently, de Melo et al (2021) used adipose-derived stem cells (ASCs) from human adipose tissue and fibroblasts from skin tissue as screening targets and used Cell-SELEX and quantitative PCR technology to screen and identify an aptamer named Apta99. The aptamer can specifically recognize ASCs and shows a low affinity for fibroblasts, which is expected to be a powerful tool in ASC purification and therapeutic applications.…”