The characterization of human stem cells for the usability in regenerative medicine is particularly based on investigations regarding\r\ntheir differentiation potential in vivo. In this regard, the chicken embryo model represents an ideal model organism. However,\r\nthe access to the chicken embryo is only achievable by windowing the eggshell resulting in limited visibility and accessibility in\r\nsubsequent experiments. On the contrary, ex ovo-culture systems avoid such negative side effects. Here, we present an improved\r\nex ovo-cultivation method enabling the embryos to survive 13 days in vitro. Optimized cultivation of chicken embryos resulted\r\nin a normal development regarding their size and weight. Our ex ovo-approach closely resembles the development of chicken\r\nembryos in ovo, as demonstrated by properly developed nervous system, bones, and cartilage at expected time points. Finally, we\r\ninvestigated the usability of our method for trans-species transplantation of adult stemcells by injecting human neural crest-derived\r\nstem cells into late Hamburger and Hamilton stages (HH26ââ?¬â??HH28/E5ââ?¬â?E6) of ex ovo-incubated embryos. We demonstrated the\r\nintegration of human cells allowing experimentally easy investigation of the differentiation potential in the proper developmental\r\ncontext. Taken together, this ex ovo-method supports the prolonged cultivation of properly developing chicken embryos enabling\r\nintegration studies of xenografted mammalian stem cells at late developmental stages.
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