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. 2017 Sep;26(9):1582-1595.
doi: 10.1177/0963689717723015.

Porcine Neural Progenitor Cells Derived from Tissue at Different Gestational Ages Can Be Distinguished by Global Transcriptome

Jing Yang  1   2 , Steven Menges  1   2 , Ping Gu  1   2   3 , Ronald Tongbai  1   4 , Melissa Samuel  5 , Randall S Prather  5 , Henry Klassen  1   2
Affiliations

Porcine Neural Progenitor Cells Derived from Tissue at Different Gestational Ages Can Be Distinguished by Global Transcriptome

Jing Yang et al. Cell Transplant. 2017 Sep.

Abstract

The impact of gestational age on mammalian neural progenitor cells is potentially important for both an understanding of neural development and the selection of donor cells for novel cell-based treatment strategies. In terms of the latter, it can be problematic to rely entirely on rodent models in which the gestational period is significantly shorter and the brain much smaller than is the case in humans. Here, we analyzed pig brain progenitor cells (pBPCs) harvested at 2 different gestational ages (E45 and E60) using gene expression profiles, obtained by microarray analysis and quantitative polymerase chain reaction (qPCR), across time in culture. Comparison of the global transcriptome of pBPCs from age-matched transgenic green flourescent protein (GFP)-expressing fetuses versus non- GFP-expressing fetuses did not reveal significant differences between the 2 cell types, whereas comparison between E45 and E60 pBPCs did show separation between the data sets by principle component analysis. Further examination by qPCR showed evidence of relative downregulation of proliferation markers and upregulation of glial markers in the gestationally older (E60) cells. Additional comparisons were made. This study provides evidence of age-related changes in the gene expression of cultured fetal porcine neural progenitors that are potentially relevant to the role of these cells during development and as donor cells for transplantation studies.

Keywords: brain; gene expression profile; microarray; stem cell; transgenic swine.

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Conflict of interest statement

Declaration of Conflicting Interests: The author(s) declared the following potential conflicts of interest with respect to the research, authorship, and/or publication of this article: Henry Klassen and Jing Yang have an equity interest in jCyte, Inc., a company that may potentially benefit from the research results presented. They also serve on the company’s board. The terms of this arrangement have been reviewed and approved by the University of California, Irvine, in accordance with its conflict of interest policies.

Figures

Fig. 1.
Fig. 1.
Principal component analysis of E45 GFP transgenic pBPCs versus E45 nontransgenic pBPCs, in both SM and UL. E45 GFP-transgenic pBPC is in green, and E45 nontransgenic pBPC is in red. SM, standard medium; pBPCs, pig brain progenitor cells.
Fig. 2.
Fig. 2.
(A) Principal component analysis of E60 nontransgenic pBPC versus E45 nontransgenic pBPC, both in SM. E60 pBPC is in blue, and E45 pBPC is in red. (B) Quantitative real-time PCR analysis of select progenitor and differentiation markers between the E60 pBPCs in SM and E45 pBPCs in SM. Data are normalized to E45. Values greater than 1 indicate higher expression in E60 cells and values less than 1 indicate higher expression in E45 cells. The reference line at Relative Quantification (RQ) = 1 indicates no fold change difference. *P < 0.05 for E60 versus E45. SM, standard medium; pBPC, pig brain progenitor cells; PCR, polymerase chain reaction; pBPCs, pig brain progenitor cells.
Fig. 3.
Fig. 3.
Image taken of E45 pBPC cells in SM (A) and UL (B) taken after 42 d in culture. Higher magnification of E45 pBPC cells in SM (C) and UL (D) is also shown. SM, standard medium; pBPC, pig brain progenitor cells.
Fig. 4.
Fig. 4.
(A) Principal component analysis of E45 pBPCs in UL versus E45 pBPCs in SM. E45 pBPC in UL is in blue, and E45 pBPC in SM is in green. (B) Quantitative real-time PCR analysis of select progenitor and differentiation markers between the E45 pBPCs in UL and SM. Data are normalized to E45 in SM. Values greater than 1 indicate higher expression in E45 UL-treated cells, and values below 1 indicate higher expression in E45 SM-treated cells. The reference line at Relative Quantification (RQ) = 1 indicates no fold change difference. *P < 0.05 for E45 UL versus SM. SM, standard medium; pBPC, pig brain progenitor cells; PCR, polymerase chain reaction; pBPCs, pig brain progenitor cells.
Fig. 5.
Fig. 5.
CLICK clustering analysis of E45 pBPCs in SM (A) and UL (B) with time. SM, standard medium; pBPCs, pig brain progenitor cells.
Fig. 6.
Fig. 6.
CLICK clustering analysis of E60 pBPCs with time. pBPCs, pig brain progenitor cells.
See this image and copyright information in PMC

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