Open Access

ARTICLE

Transplanted choroidal plexus epithelial cells can integrate with organotypic spinal cord slices into a new system

by JINGJIE LIU^1,#, XIAOYAN DING^2,#, LI XIANG¹, SHENGLI HUANG^3,*

1 Department of Neurology, The Second Affiliated Hospital, College of Medicine, Xi’an Jiaotong University, Xi’an, 710004, China
2 Department of Ophthalmology, Xi’an No. 3 Hospital, Xi’an, 710018, China
3 Department of Orthopedics, The Second Affiliated Hospital, College of Medicine, Xi’an Jiaotong University, Xi’an, 710004, China

* Address correspondence to: Shengli Huang,
# These authors contributed equally to this work

(This article belongs to the Special Issue: Cell-Based Regenerative Therapies)

BIOCELL 2022, 46(6), 1537-1544. https://doi.org/10.32604/biocell.2022.018441

Received 25 July 2021; Accepted 19 October 2021; Issue published 07 February 2022

Abstract

This study aimed to evaluate the integration of transplanted choroidal plexus epithelial cells with organotypic spinal cord slices. Organotypic spinal cord slices, normally cultured for 6 days, were divided into control group (Ctrl) and transplanted group (T). The choroidal plexus epithelial cells were dissociated and primary cultured (C group). The choroidal plexus epithelial cells cultured for 6–7 days were labeled by 1,1’-dioctadecyl-3,3,3’,3’-tetramethyl-indocarbocyanineperchlorate (CM-Dil), and were identified by transthyretin (TTR) in immunocytochemistry. They were adjusted to the density of 0.5–1 × 10⁷/ml, then 2 μl cells suspension were transplanted to the spinal cord slices in the T group. The same amount of basal medium was dripped on the spinal cord slices in the Ctrl group. After 14 days of transplantation, the differentiations into neurons and astrocytes, and the synapses were identified by immunofluorescence histochemistry. At the same time, the ratios of cell differentiations and synapses in new system, and the changes of MAPK signaling pathway were tested by western blotting. The choroid plexus epithelial cells were well labeled by CM-Dil and were immune-stained by TTR in immunocytochemistry. The choroid plexus epithelial cells bodies were small when transplanted on the spinal cord slices, but big when transplanted on the polyester membrane inserts. The transplanted cells could differentiate into astrocytes, and possibly differentiate into neurons, and there were a large number of synaptophysin positive vesicles between transplanted cells and organotypic spinal cord slices in immunofluorescence histochemistry. The levels of GFAP, TUB-III and synaptophysin in the T group were higher than which in the Ctrl and C groups in western blotting (P < 0.05). And the ratios of p-JNK/JNK and p-P38/P38 in the T group were significantly lower than which in the Ctrl and C groups (P < 0.05). But the ratio of p-ERK/ERK in the three groups was of no significant difference. The transplanted choroidal plexus epithelial cells can integrate with organotypic spinal cord slices into a new system.

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