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Preprints

Enhance the Osteogenic Differentiation of Human Pluripotent Stem Cells Through the Addition of Osteogenesis Peptide During Cell Type-based Periods

Song Y, Li H, Feng F, Shi J, Li J, Wang L, Liao L, Xie B, Ma S, Li S, Zhang Y, Liu B, Yang Y, Zhou P.
Preprint from
Research Square
16 March 2022
PPR
PPR469595
Abstract

Background:

The in vitro osteogenic differentiation of human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) have been one of the hot topics in bone tissue engineering. Moreover, supplement of osteogenesis chemical compounds is commonly applied to improve the differentiation efficiency. However, the differentiation process of hPSCs is much more complex than adult stem cells, and the unknown effect of material addition at different stages hamper the establish of step-wise induction systems for these human pluripotent stem cells (hPSCs). Recently, a bone forming peptide-1 (BFP-1) that derived from bone morphogenetic protein-7 (BMP-7) has confirmed its excellent performance in promoting the osteogenic induction of human stem cells mainly human mesenchymal stem cells.

Methods:

: In this study, we cultured hESCs and hiPSCs in the osteogenic induction medium for 28 days, and added BFP-1 at varying weeks. After differentiation for varying days (0, 7, 14, 21 and 28), we investigated the expression of related gene/protein markers and the differentiation efficiency by RT-PCR, immunofluorescence and alizarin red staining assay. Moreover, the expression of marker genes relating to germ layers and epithelial-mesenchymal transition (EMT) was investigated at day 7, aiming to study the impact of BFP1 peptide on the lineage differentiation and osteogenic induction process of hESCs and hiPSCs.

Results:

: Biocompatible BFP1 peptide could remarkably promote the generation of mesoderm cells and mesenchymal-like cells from hiPSCs through the enhanced epithelial-mesenchymal transition, but the effect was not found in hESCs because of cell line difference. Moreover, peptide addition could apparently up-regulate the expression of marker gene/protein in hESCs, especially the differentiation efficiency was improved by determining the optimal treatment periods.

Conclusions:

: Our work has a great value in improving the in vitro osteogenic differentiation efficiency of hPSCs by adding functional osteogenesis compounds at specific stages and promoting the fundamental and clinical applications of osteoblast like cells.