Konovalova E V, Novosadova E V, Grivennikov I A, Illarioshkin S N
Research Center of Neurology, Moscow, Russia.
Institute of Molecular Genetics, the Russian Academy of Sciences, Moscow, Russia.
Bull Exp Biol Med. 2015 Oct;159(6):772-5. doi: 10.1007/s10517-015-3072-9. Epub 2015 Oct 31.
Fibroblasts isolated from skin biopsy specimens from patients with genetic forms of Parkinson's disease, carriers of mutations in LRRK2 and PARK2 genes, and from a healthy volunteer were reprogrammed using lentiviral vectors into induced pluripotent stem cells (iPSC). iPSC were differentiated into neuron-like cells using a cocktail of differentiation factors (N2, B27, and Noggin). The iPSC lines derived from patients with different mutations and from a healthy volunteer cultured under the same conditions were characterized by different proportion of neuronal precursors and differentiated neurons. Control Po2 line contained 56% precursors, while B15 line with LRRK2 gene mutation (G2019S) contained 35% precursor cells. Similar regularities were characteristic of Tr5 culture carrying compound heterozygous mutations in PARK2 gene (del202-203AG and IVS1+1G/A) and containing 4% neuronal precursors. Further comparative studies of iPSC carrying various mutations and comparison with normal human cells will help to understand the molecular pathogenesis of some genetic variants of Parkinson's disease.
从患有帕金森病遗传形式的患者、携带LRRK2和PARK2基因突变的携带者以及一名健康志愿者的皮肤活检标本中分离出的成纤维细胞，使用慢病毒载体重编程为诱导多能干细胞（iPSC）。iPSC使用分化因子混合物（N2、B27和Noggin）分化为神经元样细胞。在相同条件下培养的来自不同突变患者和健康志愿者的iPSC系，其神经元前体和分化神经元的比例不同。对照Po2系含有56%的前体，而携带LRRK2基因突变（G2019S）的B15系含有35%的前体细胞。在PARK2基因中携带复合杂合突变（del202 - 203AG和IVS1 + 1G/A）且含有4%神经元前体的Tr5培养物也有类似规律。对携带各种突变的iPSC进行进一步的比较研究，并与正常人细胞进行比较，将有助于了解帕金森病某些遗传变异的分子发病机制。
