A novel method for the isolation of subpopulations of rat adipose stem cells with different proliferation and osteogenic differentiation potentials

Abstract

Bone marrow has been the elected cell source of studies published so far concerning bone and cartilage tissue-engineering approaches. Recent studies indicate that adipose tissue presents significant advantages over bone marrow as a cell source for tissue engineering. Most of these studies report the use of adipose stem cells (ASCs) isolated by a method based on the enzymatic digestion of the adipose tissue and on the ability of stem cells to adhere to a cell culture plastic surface. Using this method, a heterogeneous population was obtained containing different cell types that have been shown to compromise the proliferation and differentiation potential of the stem cells. This paper reports the development and optimization of a new isolation method that enables purified cell populations to be obtained that exhibit higher osteogenic differentiation and/or proliferation potential. This method is based on the use of immunomagnetic beads coated with specific antibodies and it is compared with other methods described in the literature for the selection of stem cell populations, e.g. methods based on a gradient solution and enzymatic digestion. The results showed that the isolation method based on immunomagnetic beads allows distinct subpopulations of rat ASCs to be isolated, showing different stem cells marker expressions and different osteogenic differentiation potentials. Therefore, this method can be used to study niches in ASC populations and/or also allow adipose tissue to be used as a stem cell source in a more efficient manner, increasing the potential of this cell source in future clinical applications.

Human adipose tissue‐derived tenomodulin positive subpopulation of stem cells: A promising source of tendon progenitor cells

“…In previous studies, viable, homogenous and phenotypically stable subpopulations of human and rat ASCs were successfully separated from crude populations (Mihaila et al, ; Rada et al, ; Rada, Gomes, & Reis, ; Rada, Reis, & Gomes, ) using the immunomagnetic sorting (IMS) method, enabling to select subpopulations that were found to be more prone to osteogenic (Rada, Gomes, et al, ; Rada et al, ), chondrogenic (Rada, Reis, et al, ) or endothelial differentiation (Mihaila et al, ). Therefore, purification of ASC subpopulations opens new prospects for tissue engineering applications, which may take advantage of (potentially autologous) cell sources predisposed to differentiate into specific lineages.…”

“…Therefore, the main aim of this study was to identify and characterize a subpopulation of hASCs prone to tenogenic differentiation. For this purpose, hASCs subpopulations were isolated from hASCs and sorted with IMS using CD29, STRO‐1, and SSEA‐4 antibodies based on previous studies of hASCs subpopulations (Mihaila et al, ; Mihaila, Resende, Reis, Gomes, & Marques, ; Rada, Gomes, et al, ; Rada, Reis, et al, ; Rada et al, ), and tenomodulin (TNMD), a highly expressed marker in tendons and ligaments (Brandau, Meindl, Fassler, & Aszodi, ).…”

Selective Cell Recruitment and Spatially Controlled Cell Attachment on Instructive Chitosan Surfaces Functionalized with Antibodies

“…The most common and effective method currently reported for isolation of a specific cell type involves the use of magnetic beads coated with antibodies for target cells. This technique requires the attachment of magnetic beads to the cells that may be undesirable for cell-based therapeutic applications [15,16].…”

“…Compared with currently available devices found in literature the system herein proposed besides effective it is relatively low cost using a biomaterial with easy chemistry that can be later implantable [10][11][12][13][14][15][16]. The method proposed in this work allows the isolation of a target cell type from a mixed cell population with relatively high purity, decreasing cell handling where recruited cells can be used straightaway.…”

“…The most common method for cell sorting and enrichment is based on magnetic cell separation. Magnetic beads utilize a capturing surface or matrix coated with an antibody that has high affinity for a specific cell-surface marker binding target cells [15,16]. To date, few studies reported the use of a biomaterial surfaces functionalized with antibodies for cell capture.…”