The Passage 4/5 ASCs kept in complete medium was replaced with serum-free DMEM medium with 20 ng/ml of bFGF (known neuronal inducer) and 5% PAN serum replacement. The cells were maintained for 7 days in the same medium with periodical change of fresh medium for every 3 days. Later the bFGF containing medium was removed and washed with DPBS. Serum-free medium with 5% PAN serum replacement (Panexin NTA, PAN Biotech catalogue no. PO4-95700) was added to the cells. This temporal exposure (mimics in vivo exposure) of bFGF aids neuronal induction. The morphological changes were observed under Nikon Ts2FL phase contrast microscopy [22 (link)].
Martin C.A., Radhakrishnan S., Gómez Ribelles J.L., Trentz O.A., EAK N., Reddy M.S., Rela M, & Kalkura Subbaraya N. (2022). Adipose tissue derived stromal cells in a gelatin-based 3D matrix with exclusive ascorbic acid signalling emerged as a novel neural tissue engineering construct: an innovative prototype for soft tissue. Regenerative Biomaterials, 9, rbac031.
Aids AscsCells Neuronal Phase contrast microscopySerum
Corresponding Organization :
Other organizations :
Anna University, Chennai, Universitat Politècnica de València, Biomedical Research Networking Center in Bioengineering, Biomaterials and Nanomedicine, Madras Institute of Orthopaedics and Traumatology
Replacement of the Passage 4/5 ASCs kept in complete medium with serum-free DMEM medium
Addition of 20 ng/ml of bFGF (known neuronal inducer)
Addition of 5% PAN serum replacement
dependent variables
Morphological changes observed under Nikon Ts2FL phase contrast microscopy
control variables
Maintaining the cells in the same medium for 7 days with periodical change of fresh medium every 3 days
controls
Temporal exposure (mimics in vivo exposure) of bFGF to aid neuronal induction
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