Human recombinant Laminin-511 and -521 stimulates neurite outgrowth and synaptogenesis of cerebellar progenitor cells

Abstract

With increasing prevalence of neurodegenerative diseases the need for an effective treatment increases. To date pharmacological treatment is the golden standard. However, the medicines used have many adverse effects and, only work for a short time period. The best solution would be to cure the disease and thus stop the degradation of neurons and replace the already lost neurons by healthy ones. With the development of regenerative medicine, strategies to replace the lost neurons are emerging. One of the approaches includes the use of neuronal progenitor cells to differentiate and grow into functional neurons. To achieve this, a suitable in vitro environment is needed that includes an optimal use of culture medium and substrate. Laminin has been shown to be an effective substrate for neuronal cell culture purposes as it is part of the native extracellular matrix in the nervous system. However, laminin represents a family of 20 isoforms as known so far and there is still a lot unknown about the functions of each isoform on neuronal cultures. In this study the effects of eight available human recombinant laminin isoforms (i.e., -111, -121, -211, -221, -411, -421, -511 and -521) on cell attachment, differentiation, neurite outgrowth and connectivity of cerebellar progenitor cells have been assessed by means of immunofluorescence techniques. A general laminin substrate stemming from mice sarcoma (L2020) was used as a control. The results show that Laminin- 211, -221 and -411 do not possess any neurite outgrowth and differentiation abilities and thus were not included in the remaining experiments. Between the remaining substrates used no difference was found on cell attachment and neuronal differentiation. Laminin-511 and -521 increased neurite outgrowth the most and laminin-421 the least. Laminin-111 took third place. Laminin-111, -511, -521 and L2020 were included in the connectivity experiments. Laminin-521 stimulated synaptogenesis to the largest extend followed by laminin-511 and then -111. Overall, laminin-511 and -521 showed the best results in this study and performed better than a general laminin substrate that studies mostly use.