Study On The Melanocyte-like Cell Differentiated From Zebrafish Embryonic Cell In Vitro

Pigment cell provides ainimals with gorgeous and attractive appearances.It not only helps animals suit to the environment,but also plays an important role in the interactions between species and inter-species.It’s one of the factors in natural selection and evolution.Melanocyte is one of the main categories of animal pigment cells,It contains a great many of melanin granules,which appear black by absorbing and reflecting certain wavelengths of incident light.Conducting the cell biological studies of melanocytes will contribute to the further analysis of the body color patterns’ morphological diversity,the malpigmentation exploration and the development mechanisms of melanoma.In the experimental culturing and research of zebrafish embryonic cells in vitro,we have found that the zebrafish embryonic cells can be induced by the culture system containing fish serum to differentiate to melanocyte-like cells.According to this condition,we carry out research about the zebrafish embryonic cells which are capable of differentiating to produce melanocyte-like cells.And the related results are summarized as follows:1.Using 10 hours post-fertilization zebrafish embryos(10hpf,early stage of neurula)as materials,we isolate, culture and identify the zebrafish embryos,and successfully obtain the zebrafish embryonic cells finally in vitro;2.We explore whether different culture conditions such as the growth factors, zebrafish embryo extract and fish serum will impact the culturing of the zebrafish embryonic cells in vitro. And we find out that the zebrafish embryonic cells will differentiate to melanocyte-like cells with the circumstances of the fish serum;3.Morphological and structural characteristics of melanocyte-like cells,which are acquired from the addition of fish serum,show that the melanocyte-like cells’ diameters vary from 20 to 200 micrometers.And they appear black,containing many melanin granules of different sizes.Depending on different cell morphologys,the melanocyte-like cells can be divided into spherical,branched and irregular cell types;4.Combined with RT-PCR technique,we detect the expression of melanocyte stem cell marker mitfa and melanocyte cell marker tyr under the fish serum condition.Results are displayed that the zebrafish embryonic cells in the fish serum culture system can be detected of the expression of mitfa and tyr.This illustrates the zebrafish embryonic cells can be induced to produce the melanocyte stem cells and melanocytes due to the fish serum condition;5.In order to further understand the developmental status of cultured cells in the fish serum culture system,we detect expression of pluripotency markers,for instance,Nestin, Brachyury and Gata4 via immunofluorescence.These results exhibit that the culturing cells can be detected in Nestin, Brachyury and Gata4 expression,indicating that with the circumstances of the fish serum, the zebrafish embryonic cells can differentiate to melanocyte-like cells while there still exist ectodermal cells, mesodermal cells,endoderm cells and other pluripotent stem cells in this system.Above all,these studies use zebrafish embryos as materials,confirm that the zebrafish embryonic cells can differentiate to melanocyte-like cells in fish serum culture condition.And this laid the foundation for further development of melanocyte stem cells and melanocytes in the molecular regulation mechanism.