Reprogramming of blood cells into induced pluripotent stem cells as a new cell source for cartilage repair.

Yueying Li,Jiayu Chen,Qingjun Su,Yong Hai,Tie Liu,Nicholas Van Halm-Lutterodt

doi:10.1186/s13287-016-0290-7

Abstract

BackgroundAn attempt was made to reprogram peripheral blood cells into human induced pluripotent stem cell (hiPSCs) as a new cell source for cartilage repair.MethodsWe generated chondrogenic lineage from human peripheral blood via hiPSCs using an integration-free method. Peripheral blood cells were either obtained from a human blood bank or freshly collected from volunteers. After transforming peripheral blood cells into iPSCs, the newly derived iPSCs were further characterized through karyotype analysis, pluripotency gene expression and cell differentiation ability. iPSCs were differentiated through multiple steps, including embryoid body formation, hiPSC-mesenchymal stem cell (MSC)-like cell expansion, and chondrogenic induction for 21 days. Chondrocyte phenotype was then assessed by morphological, histological and biochemical analysis, as well as the chondrogenic expression.ResultshiPSCs derived from peripheral blood cells were successfully generated, and were characterized by fluorescent immunostaining of pluripotent markers and teratoma formation in vivo. Flow cytometric analysis showed that MSC markers CD73 and CD105 were present in monolayer cultured hiPSC–MSC-like cells. Both alcian blue and toluidine blue staining of hiPSC–MSC-chondrogenic pellets showed as positive. Immunohistochemistry of collagen II and X staining of the pellets were also positive. The sulfated glycosaminoglycan content was significantly increased, and the expression levels of the chondrogenic markers COL2, COL10, COL9 and AGGRECAN were significantly higher in chondrogenic pellets than in undifferentiated cells. These results indicated that peripheral blood cells could be a potential source for differentiation into chondrogenic lineage in vitro via generation of mesenchymal progenitor cells.ConclusionsThis study supports the potential applications of utilizing peripheral blood cells in generating seed cells for cartilage regenerative medicine in a patient-specific and cost-effective approach.Electronic supplementary materialThe online version of this article (doi:10.1186/s13287-016-0290-7) contains supplementary material, which is available to authorized users.

Highlights

An attempt was made to reprogram peripheral blood cells into human induced pluripotent stem cell as a new cell source for cartilage repair
The iPSC lines used in this research were positive for the pluripotent markers OCT4, NANOG, SSEA4, TRA-1-60 and TRA-1-81(Fig. 1d)
Our study showed that the generation of iPSCs from human Peripheral blood cell (PBC) may provide an alternative source of mesenchymal stem cell (MSC) for cartilage repair

Summary

Introduction

An attempt was made to reprogram peripheral blood cells into human induced pluripotent stem cell (hiPSCs) as a new cell source for cartilage repair. Cartilage tissue engineering is a promising and potential directive to provide biological replacement tissue. This may require an adequate amount of seed cells and, as it is generally known, there is a limited source of naturally occurring autologous chondrocytes. Seed cells usually obtained from an ectopic cartilage source often require additional surgery which seems to be a significant challenge in clinical practice [1]. Adult stem cells appear to be a promising alternative cell source for cartilage tissue engineering; it is undoubtedly fraught with limitations. The percentage of mesenchymal stem cells (MSCs) in the bone marrow of an adult is rather low, and the yield of stem cells from older patients has been reported to show relatively lower proliferative rates in culture.

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