br Resource table br Resource details Multiple endocrine

Resource table
Resource details Multiple endocrine neoplasia type 1 (MEN1) (also termed Wermer syndrome) is an autosomal dominant hereditary which is caused by mutations on the tumor suppressor gene MEN1. MEN1 is characterized by tumorigenesis mainly in endocrine organs including parathyroid glands, pancreatic islets, anterior pituitary gland (Thakker, 2014). Previously we reported the cryosurgery and pathological results for insulinomas of 2 patients (mother and son) in one family with MEN1 (Li et al., 2013). In the following two years, both patients suffered with adenomatosis in parathyroid and thyroid. For the reason of who sequentially had three different endocrine tumors, it is a rare but with great value resource for our society to continue research biological basis of this syndrome. Therefore, here we report that using the urine original cathepsin inhibitor from these two patients to establish MEN1 patient specific induced pluripotent stem cells (iPS) cells. In this report, the iPSC lines were generated from the 23-year-old son. Urine of the patient was collected for cells isolation and expansion. The urine cells were tested for free of mycoplasma. For reprogramming, the urine cells were transfected with episomal plasmids expressing Oct4, Sox2, Klf4 and miR-302-367 cluster. About 3weeks after transfection, many colonies with human embryonic stem cell-like morphology revealed and 2 clones were picked for expansion and frozen. One of them was selected for further characterization. The iPS cell has embryonic stem cell-like morphology with a high nuclear to cytoplasmic ratio (Fig. 1A). Integration of the exogenous transgenes in the genome of iPS cells was excluded by PCR (Fig. 1B). Mutation analysis demonstrated the iPSC line has a point mutation in exon 9 that is the exact mutation among the urine cells, iPSC and the blood cell DNA (Fig. 1C). It is a heterozygous mutation in exon-9 (c.1288 G>T, Glu430, encoding a stop codon) resulting in aberrant splicing of a defected RNA. The pluripotency markers were measured by immune-staining for Oct4, Nanog, Tra-1-60 and alkaline phosphatase (AP) (Fig. 1D, E, F), and their RNA expressions were at a comparable level with human embryonic stem cell H1 (Fig. 1G). This cell line displayed normal karyotype (Fig. 1H). Furthermore, this cell retained the potential to differentiate into three germ layers upon embryo body formation in vitro, as showed by RNA expression analysis for key genes representing different germ layers (Fig. 1I, J). Taken together, we obtained iPSC lines from a MEN1 patient who sustains a point mutant in exon 9 of Men1 gene. It is a unique cell line resource for the molecular pathology study as well as drug development of MEN1.
Materials and methods
Conflict of interest
Acknowledgements We thank all the other members in the lab of Prof. Yin-xiong Li. This work was financially supported by Haizhu District Science and Technology Plan (2011-ZD-02, 2014-CG-05), Ministry of Science and Technology 973 Program (2015CB964700) and Guangdong Province Science and Technology Plan (2016B030301007).