a novel method to derive amniotic fluid stem cells for therapeutic purposes一个新颖的方法来推导羊水干细胞用于治疗目的.pdfVIP

Phermthai et al. BMC Cell Biology 2010, 11:79 /1471-2121/11/79 METHODOLOGY ARTICLE Open Access A novel method to derive amniotic fluid stem cells for therapeutic purposes 1* 1 1 2 2 Tatsanee Phermthai , Yuparat Odglun , Suphakde Julavijitphong , Vitaya Titapant , Prakong Chuenwattana , 1 3 Chanchai Vantanasiri , Kovit Pattanapanyasat Abstract Background: Human amniotic fluid stem (hAFS) cells have become an attractive stem cell source for medical therapy due to both their ability to propagate as stem cells and the lack of ethical debate that comes with the use of embryonic stem cells. Although techniques to derive stem cells from amniotic fluid are available, the techniques have limitations for clinical uses, including a requirement of long periods of time for stem cell production, population heterogeneity and xeno-contamination from using animal antibody-coated magnetic beads. Herein we describe a novel isolation method that fits for hAFS derivation for cell-based therapy. Methods and Results: With our method, single hAFS cells generate colonies in a primary culture of amniotic fluid cells. Individual hAFS colonies are then expanded by subculturing in order to make a clonal hAFS cell line. This method allows derivation of a substantial amount of a pure stem cell population within a short period of time. Indeed, 108 cells from a clonal hAFS line can be derived in two weeks using our method, while previous techniques require two months. The resultant hAFS cells show a 2-5 times greater proliferative ability than with previous techniques and a population doubling time of 0.8 days. The hAFS cells exhibit typical hAFS cell characteristi