Placental chorionic villus explant culture in xeno-free medium: A preliminary study

摘要

Aim: to know whether placental chorionic villous-derived stem cells can grow in xenofree medium. In this study, we did explant culture of human chorionic villous in alphaminimal essentialmedium(MEM) supplemented with humanAB serum. Experimental: The chorionic villouswas washed briefly, and minced in complete alpha MEM medium, which contained penicillin/streptomycin, amphotericin B, 1%L-glutamine and 10%human AB serum. Two to three explants (diameter 2-3 mm) was placed in each well of a 12 well plate, and several drops of the complete medium were added. Further, the plate was incubated and when the explant attached to the base of the well, 0.5 mL fresh medium was added. Further, medium changes were done every 2-3 days. Observation was done daily to detect cell growth, and cell morphology. The day when fibroblastic cell growth and appearance of various morphologies of cells in each well was first detected was noted and tabulated. Results: Two types of cells were grown, i.e. fibroblastic and non-fibroblastic cells. The fibroblastic cells began to grow and attach on day-3 to day-5. The various morphologies of cells were apparent on day-5 to day-7. Long-term explant culture, more than four weeks, showed that hematopoietic stem cell derived osteoclast-like multinuclear non-fibroblastic cells were prominent and dominated the culture field. Conclusion: Placental chorionic villous explant culture can grow well in 10% human AB serum containing alpha MEM, but when washing is inadequate, the growing cells were dominated by osteoclastlike cells.