Enrichment of mouse germline stem cells on polyelectrolyte multilayers

• Main Authors: Ai-Yun Shih, 施藹芸
• Other Authors: 蔡偉博
• Format: Others
• Language: en_US
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/87603900638788357177

碩士 === 國立臺灣大學 === 化學工程學研究所 === 98 === Polyelectrolyte multilayers (PEMs) by layer-by-layer deposition have recently been applied on approach to stem cells cultivation. This simple and versatile technique provides wide potential for variation of surface characters by applying diverse materials or insertion of biomolecules. In this study, enrichment of mouse germline stem cells (GSCs), which have potential for induction into pluripotent stem cells, achieved by variety of PEMs was investigated. First, GSCs were cultured on PEMs of polyelectrolyte poly(allylamine hydrochloride)/poly(acrylic acid) (PAH/PAA) assembled at either pH 2.0 or 6.5 in comparison with laminin and TCPS used as controls. An adjustment of deposited layer pairs from 3 to 20.5 bilayers was following proceeded on pH 6.5 films. We found that GSCs formed a greater number of loosely attached spherical colonies on pH 6.5 films than pH 2.0 or conventional culture. The number and size of colonies varied with the numbers of deposited PEM layers. Alkaline phosphatase (AP) activity and expression of Octamer-4 (Oct4) in these colonies indicated their potential as pluripotent stem cells. In the meanwhile, the ability of neuron-lineage differentiation suggested their great potential as well. In the second part of this study, the potential of immobilizing laminin-derived peptides on PEMs and altering assembled polyelectrolyte to promote the self-renewal of GSCs were assessed. SIKVAV, YIGSR and RGD were conjugated to PAH and incorporated on a 3 bilayers of PAH/PAA films assembled at pH 6.5. Enhancement of undifferentiated GSCs compared with pristine PEMs was found on the SIKVAV-conjugated films. Three bilayers of PEMs fabricated by natural polymers such as collagen (COL), gelatin (GEL), chitosan (CHI), alginate (ALG) and hyaluronic acid (HA) were utilized as substrates for GSC culture as well. However, colonies grew poorly on these films. Together, our results showed pH 6.5 PAH/PAA films incorporated with SIKVAV peptide provide a versatile substratum for self-renewal of GSC-colonies.