XENO-FREE AND A FEEDER FREE SELF-RENEWAL EXTRACELLULAR MATRIX FOR LONG-TERM MAINTENANCE OF UNDIFFERENTIATED HUMAN PLURIPOTENT STEM CELLS AND METHOD OF SYNTHESIZING THE SAME

摘要

The embodiments herein provide a xeno-free and a feeder free self-renewal extracellular matrix for long-term maintenance of undifferentiated Human Embryonic Stem Cells (hESCs) and undifferentiated Human Pluripotent Stem Cells (hPSCs) and a method of synthesizing the same. The extracellular matrix includes a conditioned medium comprising a neurobasal medium, a DMEM/F12 medium, a plurality of additives and a plurality of feeder cells. The plurality of additives are L-glutamine, β-mercaptoethanol, nonessential amino acids and insulin-transferrin-selenite. The feeder cells are Human Dermal Fibroblasts (HDFs). The conditioned medium may be added with a Rho-associated Coiled Kinase (ROCK) inhibitor Y-27632. The extracellular matrix is in the form of a gel. The method comprises preparing a conditioned medium and incubating the conditioned medium for 24 hours or 72 hours at 37° C. The method comprises coating a plate with the prepared conditioned medium for 5 min or 15 min.