Induced pluripotent stem cell reprogramming by integration-free sendai virus vectors from peripheral blood of patients with craniometaphyseal dysplasia

I. Ping Chen, Keiichi Fukuda, Noemi Fusaki, Akihiro Iida, Mamoru Hasegawa, Alexander Lichtler, Ernst J. Reichenberger

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Studies of rare genetic bone disorders are often limited due to unavailability of tissue specimens and the lack of animal models fully replicating phenotypic features. Craniometaphyseal dysplasia (CMD) is a rare monogenic disorder characterized by hyperostosis of craniofacial bones concurrent with abnormal shape of long bones. Mutations for autosomal dominant CMD have been identified in the ANK gene (ANKH). Here we describe a simple and efficient method to reprogram adherent cells cultured from peripheral blood to human induced pluripotent stem cells (hiPSCs) from eight CMD patients and five healthy controls. Peripheral blood mononuclear cells (PBMCs) were separated from 5-7 mL of whole blood by Ficoll gradient, expanded in the presence of cytokines and transduced with Sendai virus (SeV) vectors encoding OCT3/4, SOX2, KLF4, and c-MYC. SeV vector, a cytoplasmic RNA vector, is lost from host cells after propagation for 10-13 passages. These hiPSCs express stem cell markers, have normal karyotypes, and are capable of forming embryoid bodies in vitro as well as teratomas in vivo. Further differentiation of these patient-specific iPSCs into osteoblasts and osteoclasts can provide a useful tool to study the effects CMD mutations on bone, and this approach can be applied for disease modeling of other rare genetic musculoskeletal disorders.

Original languageEnglish
Pages (from-to)503-513
Number of pages11
JournalCellular Reprogramming
Volume15
Issue number6
DOIs
Publication statusPublished - 2013 Dec 1

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Sendai virus
Induced Pluripotent Stem Cells
Bone and Bones
Inborn Genetic Diseases
Hyperostosis
Embryoid Bodies
Ficoll
Mutation
Teratoma
Osteoclasts
Osteoblasts
Karyotype
Cultured Cells
Blood Cells
Stem Cells
Animal Models
RNA
Cytokines
Schwartz-Lelek syndrome
Cellular Reprogramming

ASJC Scopus subject areas

  • Biotechnology
  • Developmental Biology
  • Cell Biology

Cite this

Induced pluripotent stem cell reprogramming by integration-free sendai virus vectors from peripheral blood of patients with craniometaphyseal dysplasia. / Chen, I. Ping; Fukuda, Keiichi; Fusaki, Noemi; Iida, Akihiro; Hasegawa, Mamoru; Lichtler, Alexander; Reichenberger, Ernst J.

In: Cellular Reprogramming, Vol. 15, No. 6, 01.12.2013, p. 503-513.

Research output: Contribution to journalArticle

Chen, I. Ping ; Fukuda, Keiichi ; Fusaki, Noemi ; Iida, Akihiro ; Hasegawa, Mamoru ; Lichtler, Alexander ; Reichenberger, Ernst J. / Induced pluripotent stem cell reprogramming by integration-free sendai virus vectors from peripheral blood of patients with craniometaphyseal dysplasia. In: Cellular Reprogramming. 2013 ; Vol. 15, No. 6. pp. 503-513.
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