Conditioned media trans-differentiate mature fibroblasts into pancreatic beta-like cells.
OBJECTIVE
The study was carried out to evaluate the role of preconditioning strategies on the trans-differentiation of mature fibroblasts (NIH3T3 cells) into insulin producing β-cells.


METHODS
The NIH3T3 cells were treated with dexamethasone (5μM) and pancreatic extract (0.05 and 0.4mg/mL) separately or in combination. The treated cells were analyzed for the morphological changes, and expression of pancreatic genes and proteins by phase contrast microscopy, RT-PCR and flow cytometry/immunocytochemistry, respectively.


RESULTS
Treatment of mature fibroblasts with different combinations of dexamethasone and pancreatic extract in the form of conditioned media resulted in comparable morphological changes and expression of certain pancreatic genes and proteins; however, their expression varied with each treatment. Most prominent effect was observed in case of combined treatment which resulted in significant increase (p<0.001) in gene expression levels of insulin, MafA, and Ngn3. Variable pattern was observed in insulin, MafA, Ngn3 and Sca1 expressions at the protein level.


CONCLUSIONS
It is concluded from this study that preconditioning of NIH3T3 cells with conditioned media containing different combinations of dexamethasone and pancreatic extract can induce trans-differentiation of these cells into pancreatic β-like cells. The conditioned media however, need to be optimized. The study may offer the possibility of improved regeneration of mature cell type that could serve as a future therapeutic option for diabetes.
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan.
Shumaila
S
Usman
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan.
Irfan
I
Khan
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan; Dow University of Health Sciences, Ojha Campus, Gulzar-e-Hijri,Suparco Road, KDA Scheme-33, Karachi, Pakistan.
Nadia
N
Naeem
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan.
Hana'a
H
Iqbal
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan; Department of Physiology, University of Karachi, Karachi-75270, Pakistan.
Anwar
A
Ali
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan.
Sehrish
S
Usman
Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi-75270, Pakistan. Electronic address: asmat.salim@iccs.edu.
Asmat
A
Salim
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D017077:Culture Media, Conditioned / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects*; D006801:Humans; D050417:Insulin-Secreting Cells / Q000166:cytology* / Q000187:drug effects; D051379:Mice; D041681:NIH 3T3 Cells; D010179:Pancreas / Q000166:cytology; D010184:Pancreatic Extracts / Q000235:genetics / Q000494:pharmacology; D016133:Polymerase Chain Reaction
D017077:Culture Media, Conditioned; D010184:Pancreatic Extracts; D003907:Dexamethasone
Dexamethasone; Direct differentiation; Pancreatic extract; Reactive oxygen species; Type 1 diabetes