Cultivation And Differentiation Into Neurons Of ADSCs From Rat As Well As The Expression Of BDNF Gene And Receptor Of TrkB

Objectives: To probe that whether can ADSCs be isolated fromnormal rat adipose tissue, furthermore to make inspections into themorphology, growth pattern, and adhersion rate as well as the growthcurve, ect. Attempt to induce ADSCs towards neurogenetic differentiationand derive the outcome via ameliorated induce agent in Bone MarrowStem Cells (BMSCs) induction. Through quantify expression of Nestin ofADSCs in preinduction stage, to find the underlying relationship amidADSCs differentiation and NSCs in this intermediate period. At lengthcompare the level of expression of BDNF gene before and induction,meanwhile including its receptor trkB. The terminal aim is to get a kindof adult stem cells confirmed can self-duplicated, successive passage,and transform to neurocytes, with the ability to carry out neurocytes'functions.Methods: In first segment, to get fat tissue from inguinal andpostperitoneal of rat, then to isolate ADSCs by mechanical incision anddigestive way. Innoculate them into 15%FBS BMDM media, and recordshape under contrast phase microscope. On confluence, to passage them.Put the third passage into gauge for adhersion rate and growth curve byMTT, also identify the cells by stem cells common marker CD44 byimmunofluorescence stain. In second segment, to induce ADSCs toneurocytes with the two stages strategy, after differentiation use anotherimmunofluorescence stain to make sure If cells express the specificmarker NSE, GFAP for induction results. And morphology change-andNestin dynamic expression within preinduction stage are found respectively by contrast phase microscope and immunohistochemistry.Last segment, by RT—PCR and immunohistochemistry to detect theinfluence from induction over expression of gene BDNF and its receptorrespectively. For distinction, result should be analysis by tool SPSS 11.0.Result: Indeed, we can reap ADSCs from rat fat tissue, and we frogthem show a fusiform shape after stretching and rapid adhersion rate.They also can passage seeming infinitely keeping considerable magnitudeamount. Expression of CD44 that can designate a stem cells is drasticlypositive in ADSCs. After a corporate induction with NPMM and BME,most of induced cells express NSE, as a specific marker for neuron whichalso figure the neuron-shape of these cells. In the course of differentiation,Nestin gradually descend in expression, as deprecates a intimate bondbetween differentiating cells. We, luckily have detect a propelling effectin expression of gene BDNF from induction, and BDNF's receptor trkBparallel the ascent of gene BDNF.Conclusion: There are robust abilities for ADSCs in duplication invitro, and differentiation into neuron. Distinctive change can be known inmorphology, surface marker and function, which can not but underlysADSCs as a optimum adult stem cells in therapy CNS traumatic ordegeneration diseases via substitution and carrier roles at least.