Purification of functional human ES and iPSC-derived midbrain dopaminergic progenitors using LRTM1.

Bumpei Samata,Daisuke Doi,Akira Watanabe,Yuichi Ono,Jun Takahashi,Tetsuhiro Kikuchi,Kaneyasu Nishimura,Yoshimasa Sakamoto,Jungo Kakuta

doi:10.1038/ncomms13097

Bumpei Samata, Daisuke Doi + Show 7 more

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https://doi.org/10.1038/ncomms13097

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Journal: Nature communications	Publication Date: Oct 14, 2016
Citations: 84	License type: CC BY 4.0

Affiliation: Kyoto University, KAN Research Institute

Abstract

Human induced pluripotent stem cells (iPSCs) can provide a promising source of midbrain dopaminergic (mDA) neurons for cell replacement therapy for Parkinson's disease (PD). However, iPSC-derived donor cells inevitably contain tumorigenic or inappropriate cells. To eliminate these unwanted cells, cell sorting using antibodies for specific markers such as CORIN or ALCAM has been developed, but neither marker is specific for ventral midbrain. Here we employ a double selection strategy for cells expressing both CORIN and LMX1A::GFP, and report a cell surface marker to enrich mDA progenitors, LRTM1. When transplanted into 6-OHDA-lesioned rats, human iPSC-derived LRTM1+ cells survive and differentiate into mDA neurons in vivo, resulting in a significant improvement in motor behaviour without tumour formation. In addition, there was marked survival of mDA neurons following transplantation of LRTM1+ cells into the brain of an MPTP-treated monkey. Thus, LRTM1 may provide a tool for efficient and safe cell therapy for PD patients.

Highlights

Human induced pluripotent stem cells can provide a promising source of midbrain dopaminergic neurons for cell replacement therapy for Parkinson’s disease (PD)
During early development of mouse brain, LMX1A is expressed in midbrain[14], whereas CORIN is expressed in FP9
Double-labelled immunostaining revealed that CORIN þ LMX1A::GFP þ cells gave rise to midbrain dopaminergic (mDA) neurons, which expressed TH, NURR1 and dopamine transporter (DAT), more frequently than unsorted cells (Fig. 1g–o). These results indicate that mDA progenitors were enriched in the CORIN þ LMX1A::GFP þ population

Summary

Introduction

Human induced pluripotent stem cells (iPSCs) can provide a promising source of midbrain dopaminergic (mDA) neurons for cell replacement therapy for Parkinson’s disease (PD). IPSC-derived donor cells inevitably contain tumorigenic or inappropriate cells To eliminate these unwanted cells, cell sorting using antibodies for specific markers such as CORIN or ALCAM has been developed, but neither marker is specific for ventral midbrain. When transplanted into 6-OHDA-lesioned rats, human iPSC-derived LRTM1 þ cells survive and differentiate into mDA neurons in vivo, resulting in a significant improvement in motor behaviour without tumour formation. To overcome this problem, one possible strategy is to combine a refined differentiation protocol that induces mesencephalon with FACS using anti-CORIN antibody[9]. HiPSC-derived LRTM1 þ cells survived well in a short-term primate study without overgrowth

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