P-797 A novel method for establishing human embryonic stem cells independent of feeder cells

AbstractStudy questionIs there a efficient establishing method of human embryonic stem cells directly from the human blastocysts independent of feeder cells?Summary answerWe established a novel method of generating human embryonic stem cells directly from human blastocysts independent of feeder layer cells.What is known alreadyEstablishing embryonic stem cells lines mainly needed to coculture ICM clumps with feeder cells (like mouse or human fibroblasts), this brought in potential heterogeneous pollution. Although there had be some reports about generating human ESCs independent of feeder cells, but the efficiency was low and conditioned medium were unstable and also had the biological contamination.Study design, size, durationWe used ten day5/6 donated human blastocysts from our reproductive center, most of them were genetically diseased embryos with abnormal PGT diagnosis. After establishing ESCs procedure, all the cell lines were identified with pluripotency and differentiation potential tests. The success rate of system was calculated and compared with the conventional methods.Participants/materials, setting, methodsIn brief, ICM clumps were separated mechanically by using a micromanipulation system,and then transferred to a 30ul mTESR plus culture media drop pretreated with the geltrex (1:100 dilution) matrix and oxygen concentration was 5%. When cells attached and migrated,we also used laser to destroy the remaining trophoblast cells. About 10 days,the typical ES clone ...
Source: Human Reproduction - Category: Reproduction Medicine Source Type: research