In vitro differentiation of human bone marrow stromal cells into neural precursor cells using small molecules

Abeer Sallam; Thangirala Sudha; Noureldien H E Darwish; Samar Eghotny; Abeer E-Dief; Passainte S Hassaan; Shaker A Mousa

doi:10.1016/j.jneumeth.2021.109340

In vitro differentiation of human bone marrow stromal cells into neural precursor cells using small molecules

J Neurosci Methods. 2021 Nov 1:363:109340. doi: 10.1016/j.jneumeth.2021.109340. Epub 2021 Aug 28.

Authors

Abeer Sallam¹, Thangirala Sudha², Noureldien H E Darwish³, Samar Eghotny⁴, Abeer E-Dief⁴, Passainte S Hassaan⁴, Shaker A Mousa⁵

Affiliations

¹ Department of Medical Physiology, Faculty of Medicine, Alexandria University, Alexandria, Egypt; Center of Excellence for Research in Regenerative Medicine and its Applications, Faculty of Medicine, Alexandria University, Alexandria, Egypt.
² The Pharmaceutical Research Institute, Albany College of Pharmacy and Health Sciences, Rensselaer, NY, USA.
³ The Pharmaceutical Research Institute, Albany College of Pharmacy and Health Sciences, Rensselaer, NY, USA; Hematology Unit, Clinical Pathology Department, Faculty of Medicine, Mansoura University, Mansoura, Egypt.
⁴ Department of Medical Physiology, Faculty of Medicine, Alexandria University, Alexandria, Egypt.
⁵ The Pharmaceutical Research Institute, Albany College of Pharmacy and Health Sciences, Rensselaer, NY, USA. Electronic address: shaker.mousa@acphs.edu.

PMID: 34461154
DOI: 10.1016/j.jneumeth.2021.109340

Abstract

Background: Neurogenic differentiation of human marrow stromal stem cells (hMSCs) into neural precursor cells (NPCs) offers new hope in many neurological diseases. Stromal cells can be differentiated into NPCs using small molecules acting as chemical inducers. The aim of this study is to formulate an efficient, direct, fast and safe protocol to differentiate hMSCs into NPCs using different inducers: b-mercaptoethanol (BME), triiodothyronine (T3), and curcumin (CUR). NEW METHOD: hMSCs were subjected to either 1 mM BME, 0.5 µM T3, or 5 µM CUR. Neurogenic differentiation was determined by assessing the protein expression of PAX6, SOX2, DLX2, and GAP-43 with flow cytometry and immunofluorescence, along with Nissl staining of differentiated cells.

Results and comparison with existing method: It was revealed that T3 and CUR are 70-80% better than BME in terms of efficiency and safety, and surprisingly BME was a good promoting factor for cell preconditioning with limited effects on neural trans-differentiation related to its toxic effects on cell viability.

Conclusion: Reprogramming of bone marrow stromal cells into neural cells gives hope for treating different neurological disorders. Our study shows that T3 and CUR were effective in generation of NPCs from hMSCs with preservation of cell viability. BME was a good promoting factor for cell preconditioning with limited effects on neural transdifferentiation related to its toxic effects on cell viability.

Keywords: Curcumin (CUR); DLX2; GAP-43; Human marrow stromal stem cells (hMSCs); Neural precursor cells (NPCs); Neuronal cell reprogramming; PAX6; SOX2; Small molecules; Triiodothyronine (T3).

MeSH terms

Bone Marrow Cells
Cell Differentiation
Humans
Mesenchymal Stem Cells*
Neural Stem Cells*
Neurons