NDLI: Generation of induced pluripotent stem cells from buffalo (Bubalus bubalis) fetal fibroblasts with buffalo defined factors.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Deng, Yanfei Liu, Qingyou Luo, Chan Chen, Shibei Li, Xiangping Wang, Caizhu Liu, Zhenzhen Lei, Xiaocan Zhang, Huina Sun, Hongliang Lu, Fenghua Jiang, Jianrong Shi, Deshun
Description	Author Affiliation: Deng Y ( State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, Guangxi University , Nanning, People's Republic of China .)
Abstract	Ectopically, expression of defined factors could reprogram mammalian somatic cells into induced pluripotent stem cells (iPSCs), which initiates a new strategy to obtain pluripotent stem cell lines. Attempts have been made to generate buffalo pluripotent stem cells by culturing primary germ cells or inner cell mass, but the efficiency is extremely low. Here, we report a successful method to reprogram buffalo fetal fibroblasts (BFFs) into pluripotent stem cells [buffalo induced pluripotent stem cell (biPSCs)] by transduction of buffalo defined factors (Oct4, Sox2, Klf4, and c-Myc) using retroviral vectors. The established biPSCs displayed typical morphological characteristics of pluripotent stem cells, normal karyotype, positive staining of alkaline phosphatase, and expressed pluripotent markers including Oct4, Sox2, Nanog, Lin28, E-Cadherin, SSEA-1, SSEA-4, TRA-1-81, STAT3, and FOXD3. They could form embryoid bodies (EBs) in vitro and teratomas after injecting into the nude BALB/C mice, and 3 germ layers were identified in the EBs and teratomas. Methylation assay revealed that the promoters of Oct4 and Nanog were hypomethylated in biPSCs compared with BFFs and pre-biPSCs, while the promoters of Sox2 and E-Cadherin were hypomethylated in both BFFs and biPSCs. Further, inhibiting p53 expression by coexpression of SV40 large T antigen and buffalo defined factors in BFFs or treating BFFs with p53 inhibitor pifithrin-a (PFT) could increase the efficiency of biPSCs generation up to 3-fold, and nuclear transfer embryos reconstructed with biPSCs could develop to blastocysts. These results indicate that BFFs can be reprogrammed into biPSCs by buffalo defined factors, and the generation efficiency of biPSCs can be increased by inhibition of p53 expression. These efforts will provide a feasible approach for investigating buffalo stem cell signal pathways, establishing buffalo stem cell lines, and producing genetic modification buffaloes in the future.
File Format	HTM / HTML
ISSN	15473287
Issue Number	13
Volume Number	21
Journal	Stem Cells and Development
e-ISSN	15578534
Language	English
Publisher	Mary Ann Liebert, Inc.
Publisher Date	2012-09-01
Publisher Place	United States
Access Restriction	Subscribed
Subject Keyword	Discipline Cell Biology Discipline Hematology Buffaloes Embryology Fetus Cytology Fibroblasts Induced Pluripotent Stem Cells Animals Antigens, Polyomavirus Transforming Genetics Metabolism Benzothiazoles Pharmacology Biological Markers Blastocyst Cadherins Cell Differentiation Cell Proliferation Cells, Cultured Dna Methylation Embryoid Bodies Epigenesis, Genetic Genetic Vectors Germ Layers Kruppel-like Transcription Factors Mice Mice, Inbred Balb C Mice, Nude Nuclear Transfer Techniques Octamer Transcription Factor-3 Open Reading Frames Promoter Regions, Genetic Proto-oncogene Proteins C-myc Soxb1 Transcription Factors Simian Virus 40 Teratoma Toluene Analogs & Derivatives Tumor Suppressor Protein P53 Antagonists & Inhibitors Journal Article
Content Type	Text
Resource Type	Article
Subject	Cell Biology Developmental Biology Hematology

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