Nervous system diseases (such as Parkinson’s or post-traumatic medullar injury) are especially difficult to treat, as it is not easy to replace the parts of the neural puzzle which are damaged. The key is in developing functional neurons from in vitro-treated, cells but for this it is essential that the support on which these cells are based simulate the characteristics of the nervous system. This is what biochemist Patricia García has done, developing and validating a polymer support capable of inducing neuronal differentiation in vitro. Her PhD thesis, defended at the University of the Basque Country (UPV/EHU) and undertaken at the Tecnalia Health Unit, is entitled Development of a new polymer support with components of extracellular neural matrix for application in the in vitro differentiation of different primary cells.
Ms García has developed a polymer material which contains topographic and biochemical characteristics suitable for converting cells into functional neurones. With this purpose in mind, photolithographic techniques have been combined with the technology of extracellular neural matrixes. The photolithographic technique involves a series of treatments based on exposure to light and chemical substances, and has acted to provide the support with the desired topographical form. As regards the extracellular neural matrix, cells adhere to this natural substrate in order to, amongst other things, be guided and acquire neuronal functionality, enabling the formation of new transplantable tissue for the nervous system. Ms García simulated the properties of this matrix in the support developed.
Biocompatible and inductor
At the validation phase, this polymer material proved to be biocompatible with the nervous system. It also acts to induce the formation of neurones sufficiently developed to be able to be used therapeutically, providing the induction is carried out from neural origin cells (the trials with non-neural origin cells gave rise to immature neurones, in the best of cases). Ms García undertook validations with the PC12 rat established cell line (used in numerous trials for neuronal differentiation) and with cell cultures of different origins (primary cultures) extracted from rats.
Concretely, in the case of the validations with PC12, the biocompatibility and capacity of neuronal and dopaminergic differentiation (dopamine complies with neurotransmitter functions) of the support developed have been clearly demonstrated.
Only with those of neural origin
As regards primary cultures, Ms García carried out trials with cells from the hypocampus in the embryonic rat. These are cells not only of neural origin but with the neuronal line already determined in states of early development such as the embryonic stage. These cells differentiate more efficiently in the developed matrix than in the classic conditions of culture, obtaining mature neurones after only eight days.
The second primary culture validated in the matrix was the one consisting of neural precursor cells (cells at the stage prior to converting into neurones) of the subventricular zone (also the brain) of the neonatal rat. The results show the capacity of dopaminergic neuronal differentiation in less than 30 days of culture. Moreover, an extensive network of glial cells between the matrix and the neurones were observed, and whose function is to give trophic support to and signal the neurones in development.
The last validation trial in primary culture was undertaken with cells of non-neural origin. These are precursor cells derived from the skin of an adult rat, given that they have been already described in the literature as an important source of neural precursor cells. Nevertheless, in this case, the cells stay undifferentiated on the matrix developed, and only in a low percentage have immature neurones been obtained.
About the author
Patricia García Parra (San Sebastián, 1982) is a biochemistry graduate (UPV/EHU) and has three Masters: Molecular Biology and Biomedicine (UPV/EHU), Neuroscience and the Biology of Behaviour (University Pablo Olavide of Seville) and Management of Innovation and Technology (University of Deusto). She drew up her PhD thesis under the direction of Dr. José Iñaki Álava Marquínez (Health Unit, Tecnalia Research & Innovation-TRI) and of Dr. Fabio Cavaliere (Department of Neurosciences of Medicine at the UPV/EHU). She defended her thesis at the Department of Biochemistry and Molecular Biology at the Faculty of Science and Technology. The research was undertaken at the former Inasmet (currently belonging to TRI) with a specialisation grant from the Iñaki Goenaga Foundation, and she also collaborated with the Teófilo Hernando Institute at the Universidad Autónoma in Madrid. Currently Ms García is a post-doctoral researcher in the Instituto de Investigación Sanitaria Biodonostia (Hospital Donostia), in this Basque coastal city.English translation by: WORDLAN firstname.lastname@example.org; 615740862.