Universidad Peruana Cayetano Heredia

Isolation of adipose derived mesenchymal stem cells by using anti-CD105 VHH-magnetic beads

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dc.contributor.author Gushiken Ibañez, Eduardo
dc.contributor.author Quintana, Milagros
dc.contributor.author Agueda Perez, Luz
dc.contributor.author Herrera Velit, Rosa Patricia
dc.contributor.author Balmayor, E.R.
dc.contributor.author van Griensven, M.
dc.contributor.author Espinoza, José R.
dc.date.accessioned 2023-12-05T17:48:01Z
dc.date.available 2023-12-05T17:48:01Z
dc.date.issued 2023
dc.identifier.uri https://hdl.handle.net/20.500.12866/14640
dc.description.abstract Background: Human adipose tissue-derived stem cells (ASCs) are widely used in regenerative medicine and tissue engineering. Magnetic-activated cell sorting (MACS) with monoclonal antibodies (mAbs) that bind surface markers of stem cells such as CD105, CD73, and CD90 is currently applied for the enrichment and isolation of ASCs. Alternatives to mAbs are the variable domains of heavy chain antibodies (VHHs), which are naturally produced in camelids. We report the application of an anti-CD105 VHH conjugated to magnetic beads in the isolation of ASCs in vitro. Results: Two identical anti-CD105 VHHs (EngVHH17) were screened by phage display from a VHH cDNA library constructed from the PBMCs of an alpaca immunized with a lysate of human bladder cancer cell line (T24 cells). EngVHH17 was cloned in pET22b(+), expressed in Escherichia coli BL21 and purified in Ni-NTA chromatography resulting in a ∼15 kDa VHH antibody. EngVHH17 binds CD105 with high affinity (Kd = 3.9 × 10−10 M). The specific binding of EngVHH17 to CD105 was visualized by fluorescence inmunolabeling of phorbol-12-myristate-13-acetate (PMA)-differentiated THP1 cells using FITC labeled anti-6 × His Tag IgG1 mouse mAb. EngVHH17-magnetic beads selectively sorted human ASCs from osteoblasts in a mixed culture in vitro. The selective recovery of ASC cells using different ASC/Osteoblast ratios was higher than 85%. Conclusions: EngVHH17 coupled to magnetic beads binds CD105 expressed on the cell surface of ASCs and isolates them from osteoblasts in mixed cultures in vitro by application of an external magnetic field. EngVHH17 can be further evaluated for the isolation of MSCs by MACS. en_US
dc.language.iso eng
dc.publisher Pontificia Universidad Catolica de Valparaiso
dc.relation.ispartofseries Electronic Journal of Biotechnology
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Adipose tissue en_US
dc.subject CD105 en_US
dc.subject Magnetic beads en_US
dc.subject Mesenchymal stem cell en_US
dc.subject Osteoblasts en_US
dc.subject Regenerative medicine en_US
dc.subject Tissue engineering en_US
dc.subject VHH en_US
dc.subject.mesh Tejido Adiposo
dc.subject.mesh Endoglina
dc.subject.mesh Microesferas
dc.subject.mesh Células Madre Mesenquimatosas
dc.subject.mesh Osteoblastos
dc.subject.mesh Medicina Regenerativa
dc.subject.mesh Ingeniería de Tejidos
dc.subject.mesh Anticuerpos de Dominio Único
dc.title Isolation of adipose derived mesenchymal stem cells by using anti-CD105 VHH-magnetic beads en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1016/j.ejbt.2023.06.004
dc.relation.issn 0717-3458


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