Submission Date

5-1-2026

Document Type

Paper- Restricted to Campus Access

Department

Biochemistry & Molecular Biology

Adviser

Stephen C. Kolwicz Jr.

Committee Member

Stephen C. Kolwicz Jr.

Committee Member

Amanda Reig

Committee Member

Samantha Wilner

Department Chair

Eric Williamsen

Department Chair

Anthony Lobo

Project Description

This study investigated whether a three-stage differentiation protocol adapted from Xie et al. can drive mouse bone marrow–derived mesenchymal stem cells (BM-MSCs) toward an insulin- producing beta-like phenotype in vitro. Mouse BM-MSCs were isolated from C57BL/6N mice, expanded under standard culture conditions to passages 3–5, and then subjected to staged changes in glucose concentration and growth factors over 15 days, with outcomes assessed by morphology, dithizone (DTZ) staining for zinc-rich insulin granules, RT-PCR analysis of Glut2 expression, and LDH-based viability assays. MSC isolation and expansion were successful, yielding highly viable, spindle-shaped adherent cultures, but repeated differentiation attempts failed to generate islet-like clusters, DTZ-positive staining, or substantial Glut2 upregulation relative to mouse pancreas controls, despite overall cell viabilities near 90%. These negative results suggest that while the protocol is not broadly cytotoxic, it is ineffective at inducing a stable beta-like transcriptional and morphological phenotype in murine BM-MSCs under the conditions tested, likely reflecting species-specific differences in endocrine competence and signaling requirements. The findings highlight the need for species-tailored optimization of stage duration, factor concentrations, and molecular readouts to establish robust beta-like differentiation protocols for mouse BM-MSCs and provide a reproducible platform of viable cells for future refinement.

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