Human Adipose‐Derived Stromal Cell Isolation Methods and Use in Osteogenic and Adipogenic In Vivo Applications

Abstract

Adipose tissue represents an abundant and easily accessible source of multipotent cells, which may serve as excellent building blocks for tissue engineering. This article presents a newly described protocol for isolating adipose-derived stromal cells (ASCs) from human lipoaspirate, compared to the standard protocol for harvesting ASCs established in 2001. Human ASC isolation is performed using two methods, and resultant cells are compared through cell yield, cell viability, cell proliferation and regenerative potential. The osteogenic and adipogenic potential of ASCs isolated using both protocols are assessed in vitro and gene expression analysis is performed. The focus of this series of protocols is the regenerative potential of both cell populations in vivo. As such, the two in vivo animal models described are fat graft retention (soft tissue reconstruction) and calvarial defect healing (bone regeneration). The techniques described comprise fat grafting with cell assisted lipotransfer, and calvarial defect creation healed with cell-seeded scaffolds. © 2017 by John Wiley & Sons, Inc.

Delivery of Human Stromal Vascular Fraction Cells on Nanofibrillar Scaffolds for Treatment of Peripheral Arterial Disease

“…Freshly isolated SVF includes an abundant population of ASC, which can be identified as CD34 + /CD31 − /CD45 − cells (Tevlin et al, 2016;Brett et al, 2017a), and are mostly of pericytic or mesenchymal phenotype (Traktuev et al, 2008). Non-hematopoietic SVF cells also have a small numbers of CD31 + /CD34 + and CD31 + /CD34 − cells, which are generally identified as endothelial progenitor and mature cells, respectively (Zimmerlin et al, 2013;Polancec et al, 2019), although the latter were also referred as co-expressing CD34 + (Tallone et al, 2011).…”

Electromembrane Extraction and Mass Spectrometry for Liver Organoid Drug Metabolism Studies

“…After the formation of organoids at day 10, they were transferred into low attachment plates and cultured for another 10 days as free-floating organoids in William’s E media (Thermo Fisher Scientific), supplemented with 10 ng/mL HGF and 10 ng/mL OSM, 10 μg/mL insulin, 100 μM AAP, 0.1 μM dexamethasone, 1 μM forskolin, and 10 μM DAPT (Bio-Techne Ltd.). The iPSC line AG27 35 − 38 was differentiated using a small-molecule-driven protocol that aims to sequentially mimic in vivo liver development, resulting in hepatocyte-containing liver organoids as described by Harrison et al 39 …”

Application of tissue engineering in paediatric

“…Merecen especial interés en esta área de estudio las células madre obtenidas a partir de medula ósea y tejido adiposo (42,44), que han demostrado poseer un magnífico potencial regenerador óseo, tanto en defectos membranosos como endocondrales (40,42). Desde la descripción del tejido adiposo como potencial fuente de células multipotentes, muchos equipos trabajan en el estudio de las células estromales derivadas del tejido adiposo (ASC), así como en el desarrollo de protocolos para su obtención más óptima y su empleo en la regeneración ósea (46).…”