Gene expression changes with differentiation of cord blood stem cells to respiratory epithelial cells: a preliminary observation

Michael J Berger,Sheryl D Adams,David H Mckenna Jr,Stacey L Sprague,Barbara M Tigges,Sharon R Minnerath

doi:10.1186/scrt60

Michael J Berger, Sheryl D Adams + Show 4 more

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https://doi.org/10.1186/scrt60

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Abstract

IntroductionOwing to wide availability, low cost and avoidance of ethical concerns, umbilical cord blood (UCB) provides an attractive source of stem cells for investigational and therapeutic uses. In this study, we sought to characterize the gene expression changes as stem cells from UCB differentiate toward alveolar type II pneumocytes (ATII).MethodsControl and experimental cells were cultured in maintenance medium (mesenchymal stem cell growth medium) or differentiation medium (small airway growth medium (SAGM)), respectively, for 8 days. Total RNA was isolated from control and experimental groups for gene expression profiling and real-time polymerase chain reaction assay.ResultsAnalysis of only mixed cell lines (n = 2) with parameters including a P value of 0.01 and an intergroup gap of 2.0 yielded a set of 373 differentially expressed genes. Prominently upregulated genes included several genes associated with ATII cells and also lung cancers: ALDH3A1, VDR and CHKA. Several upregulated genes have been shown to be integral or related to ATII functioning: SGK1, HSD17B11 and LEPR. Finally, several upregulated genes appear to play a role in lung cancers, including FDXR and GP96. Downregulated genes appear to be associated with bone, muscle and central nervous system tissues as well as other widespread tissues.ConclusionsTo the best of our knowledge, this accounting of the gene expression changes associated with the differentiation of a human UCB-derived stem cell toward an ATII cell represents the first such effort. Dissecting which components of SAGM affect specific gene regulation events is warranted.

Highlights

Owing to wide availability, low cost and avoidance of ethical concerns, umbilical cord blood (UCB) provides an attractive source of stem cells for investigational and therapeutic uses
Boyse first proposed the use of human umbilical cord blood (UCB) for therapeutic reconstitution (Boyse EA et al, unpublished), and later Broxmeyer et al [1] showed that the frequency of hematopoietic progenitor cells in this graft source surpassed that found in bone marrow
Wade et al [8] documented the gene expression changes engendered by treatment of cultured fetal lung epithelial cells with dexamethasone and cyclic AMP during differentiation into alveolar type II pneumocytes (ATII) and identified a set of “hormonally responsive” genes putatively involved in this process

Summary

Introduction

Low cost and avoidance of ethical concerns, umbilical cord blood (UCB) provides an attractive source of stem cells for investigational and therapeutic uses. We sought to characterize the gene expression changes as stem cells from UCB differentiate toward alveolar type II pneumocytes (ATII). Wade et al [8] documented the gene expression changes engendered by treatment of cultured fetal lung epithelial cells with dexamethasone and cyclic AMP (cAMP) during differentiation into alveolar type II pneumocytes (ATII) and identified a set of “hormonally responsive” genes putatively involved in this process. We reported the differentiation of UCB-derived multilineage progenitor cells (MLPCs) into cells which expressed surfactant protein C (SPC) mRNA as well as SPC and which displayed morphologic features (visualized by light and transmission electron microscopy) consistent with ATII cells [9]. In general, the precise mechanisms underpinning in vitro differentiation events involving stem cells remain obscure

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