Comparison of adipose stem cells sources from various locations of rat body for their application for seeding on polymer scaffolds

Abstract

Adipose tissue yields adult adipose stem cells (ASCs) in large quantities via less-invasive methods. These cells are of interest owing to their modulating properties and paracrine activities, which can be harnessed in regenerative medicine. Many studies on the use of rat fat tissue in an autologous animal model have been conducted; however, the different locations to obtain stromal vascular fraction of rat fat depots have not been fully characterized. The purpose of the current study was to identify optimal source of ASC from various locations of rat body. Animal experiments in vitro revealed that fat depots from cervical fat are an optimal ASC source. A high ASC yield facilitates subsequent studies on autologous transplantation in rats. The secondary objective was to compare the efficiency of osteoinductive media composition and evaluate of osteogenic potential of ASCs for seeding on scaffolds for bone repair. Scaffolds were assessed in vitro, using rat adipose stem cells and threedimensional (3D) scaffolds comprising polycaprolactone (PCL) or polycaprolactone covered with tricalcium phosphate (PCL þ 5%TCP). Seeded ASCs adhere to the surface and migrate to the scaffolds. Upon staining and determining alkaline phosphatase levels, PCL þ 5%TCP scaffolds performed better than PCL scaffolds. Furthermore, growth factors such as BMP2 and FGF2 significantly increased ASC mineralization and induced osteogenesis (p < 0.05). Our results may help select and develop pre-clinical animal model for confirming the use of ASC, alone or in association with appropriate biomaterials for bone repair. ARTICLE HISTORY

Topical cell-free conditioned media harvested from adipose tissue-derived stem cells promote recovery from corneal epithelial defects caused by chemical burns

“…The expression rate of CD73, CD90, and CD105 was lower than that in human MSCs presented by the International Society of Cell therapy (ISCT) 31 . However, this has been reported a number of times in rats because of different species, isolation, culture methods, and location of fat 12 , 32 , 33 .

Figure 1 Characteristics of cultured rat adipose tissue-derived stem cells (ADSCs).

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“…Another disadvantage is the lack of previous research on limbal MSCs compared with research on MSCs derived from adipose tissue or BM. ADSCs are abundant in the human body, making them a potential material for wound healing and tissue engineering with low risks in terms of cell acquisition and easy processing 33 .…”

Scaffold vascularization method using an adipose-derived stem cell (ASC)-seeded scaffold prefabricated with a flow-through pedicle

“…Interestingly, our previous work [18] in an animal model showed that ASCs seeded into PCL and PCL + 5% TCP promoted proliferation, adhesion and osteogenic differentiation. The osteogenic assay showed that ASCs seeded on the PCL scaffold that were cultured in osteogenic medium presented significantly higher alkaline phosphatase levels during a 14-day culture period compared with ASCs cultured in the control medium (undifferentiated ASCs).…”

“…We observed that seeded ASCs adhered to the surface and migrated into scaffolds that were cultured for up to 21 days, as confirmed by DAPI staining. Moreover, the metabolic activity assessed via the MTS proliferation assay demonstrated the increase in proliferation after the 21st day of culture of ASCs on PCL [18]. However, the complete molecular process of ASC differentiation during osteogenesis and the mechanism of their cell-specific uptake remains to be fully characterized in future studies.…”

“…The ASCs from the harvested adipose tissues were collected and isolated as described previously [17,18]. Briefly, the raw adipose tissues were washed extensively with sterile phosphate-buffered saline (Life Technologies, Carlsbad, CA, USA) to eliminate debris and red blood cells.…”