Optimization of establishment of pig pluripotent cells and directly differentiation into neural lineage cells

LI Xue ZHANG Ben NIU Shu-dong WANG Yu-ge WEN Li-bo LIANG Chen QI Xiao-juan LI Yu LEI Lei*

doi:10.16098/j.issn.0529-1356.2019.01.005

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Acta Anatomica Sinica ›› 2019, Vol. 50 ›› Issue (1) : 24-28. DOI: 10.16098/j.issn.0529-1356.2019.01.005

Optimization of establishment of pig pluripotent cells and directly differentiation into neural lineage cells

LI Xue¹ ZHANG Ben² NIU Shu-dong¹WANG Yu-ge¹ WEN Li-bo¹ LIANG Chen³ QI Xiao-juan¹ LI Yu⁴ LEI Lei ^5*

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Abstract

Objective To generate the induced pig pluripotent cells (iPPCs) using the optimized induced pluripotent stem cell(iPSc) technology, and to discuss the method of directly differentiate into neural lineage cells. Methods we generated the iPPCs using the classical iPS technology in combination with valproic acid (VPA) and the methyltransferase inhibitor 5-aza-2 -deoxycytidine (5-AZA) with Oct4 retrovirus infected repetitively. Real-time PCR analysis of the expression of pluripotent related genes, immunofluorescent detection of neuron specific marker after using retinoic acid (RA) and extracellular matrix induction. Results By using this optimizing iPS technology, we successfully established a reasonable method to generate iPPCs. Real-time PCR result indicated VPA treatment significantly increased the expression of pluripotent genes, which was the same as repeated infection with Oct4 retrovirus. In addition, the iPPCs might directly differentiate into neural lineage cells after being induced with the retinoic acid and extracellular matrix. Conclusion We establishes a reasonable method to generate pig pluripotent cells, which may be a new donor cell source for human neural disease therapy.

Key words

Embryonic fibroblast / Reprogramming / Induced pluripotent cell / Neural differentiation / Retinoic acid / Immunofluorescence / Pig

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LI Xue ZHANG Ben NIU Shu-dong WANG Yu-ge WEN Li-bo LIANG Chen QI Xiao-juan LI Yu LEI Lei. Optimization of establishment of pig pluripotent cells and directly differentiation into neural lineage cells[J]. Acta Anatomica Sinica. 2019, 50(1): 24-28 https://doi.org/10.16098/j.issn.0529-1356.2019.01.005

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