Microarray analysis of microRNA expression in bone marrow-derived progenitor cells from mice with type 2 diabetes.

Bone-marrow derived vascular precursors are an important endogenous repair reservoir for vascular repair and neovascularization [1]. Therapies of stem/progenitor cells targeting on angiogenesis are considered hopeful solutions for tissue repair and regeneration. However, the dysfunction of patient-derived progenitor cells has been implicated in diabetes [2], which limited the efficacy of autologous cell therapies in the clinic [3,4]. MicroRNAs are important gene regulators whose functions remain largely unknown. In this project we reported the different microRNA expression profiles in bone marrow-derived progenitor cells from type 2 diabetic mice and their normal controls using microRNA array analysis. All microarray data are available at the Gene Expression Omnibus (GEO) at NCBI (http://www.ncbi.nlm.nih.gov/geo), under accession number GSE72616.

Direct link to deposited data

http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE72616

Experimental design, materials and methods

Animals

Male db/db (BKS.Cg-m+/+ Leprdb/J) mice and their healthy control litters (BKS.Cg-m−/− Lepdb/− lean, db/+) at the age of 10–12 weeks were purchased from the Jackson Laboratory (Bar Harbor, ME). The animals were maintained under controlled environmental condition (12 h: 12 h light/dark cycle, temperature approximately 25 °C), and provided with standard laboratory food and water ad libitum. All animal procedures were performed according to Wayne State University Institutional Animal Care and Use Committee (IACUC) guidelines.

Bone marrow-derived progenitor cell culture

The important roles of bone marrow-derived progenitor cells have been demonstrated in previous reports [1], [2]. However, they were found dysfunctional in diabetes [3], [4]. To study the microRNA profile of bone marrow-derived progenitor cells, bone marrow mononuclear cells were isolated from the tibias and femurs of mice (n = 3 each group). The cells were plated on culture flasks coated with rat plasma vitronectin (Sigma-Aldrich) and maintained in Endothelial Growth Media (EGM-2, Lonza) in 37 °C, 5% CO2. After 7 days of culture, the differentiating bone marrow-derived progenitor cells (BMPCs) as we identified in previous reports [5], [6] were used for experiments.

MicroRNA array analysis

Total RNA was extracted from db/db BMPCs and db/+ BMPCs using miRNeasy Mini Kit (Qiagen). A total of 3 μg RNA each sample was sent to perform mouse genome-wide microRNA microarray analysis using μParaflo® Biochip Technology (service provided by LC Sciences) based on the latest version of the miRBase database (Sanger miRBase Release 21). In addition, multiple control probes are included in each chip for quality controls of chip production, sample labeling and assay conditions.

Specifications
Organism/cell line/tissue	Mouse bone marrow cells
Sex	Male
Sequencer or array type	MicroRNA microarray (μParaflo® Biochip Technology)
Data format	Raw
Experimental factors	Bone marrow cells cultured in endothelial growth medium-2 for 7 days
Experimental features	Bone marrow cells were isolated from type 2 diabetes mice (db/db) and their normal control litters (db/+). The cells were cultured in endothelial growth medium-2 for 7 days. Total RNA was extracted and subjected to microRNA analysis.
Consent	Not required.
Sample source location	N/A.