Related Experiment Video
Updated: Jun 25, 2026
10:12
Differentiation of Human Pluripotent Stem Cells Into Pancreatic Beta-Cell Precursors in a 2D Culture System
Published on: December 16, 2021
Developmental stage-dependent cellular plasticity governs β-cell reprogramming efficiency in human fibroblasts
Naveen Raj1, Dhaya Shankaran Panneerselvam1, Rajkumar P Thummer1
1Laboratory for Stem Cell Engineering and Regenerative Medicine, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India.
Biochemical and Biophysical Research Communications
|June 23, 2026
Summary
Directly reprogramming fibroblasts into beta-like cells is more efficient using neonatal cells. Developmental stage significantly impacts cellular plasticity and reprogramming success for beta-cell generation.
Area of Science:
- Cellular reprogramming
- Developmental biology
- Endocrinology
Background:
- Direct reprogramming bypasses pluripotent states, reducing tumorigenesis risks.
- Generating beta-cell-like cells from fibroblasts is challenging compared to endodermal sources.
Purpose of the Study:
- To investigate the efficiency of direct reprogramming of human fibroblasts into beta-cell-like cells.
- To determine the influence of donor cell developmental stage on reprogramming efficiency.
Main Methods:
- Lentiviral-mediated reprogramming using Neurog3, Pdx1, and MafA transcription factors.
- Comparative analysis of fibroblasts from neonatal, juvenile, and adult stages.
- Validation using reporter gene analysis, microscopy, flow cytometry, RT-qPCR, and immunoblotting.
Main Results:
- Neonatal fibroblasts showed the highest reprogramming efficiency, with robust INS and MAFA expression.
- Approximately 38% of reprogrammed neonatal cells became INS-positive.
- Adult fibroblasts exhibited significantly lower reprogramming responsiveness.
- Juvenile fibroblasts showed a more heterogeneous endocrine-like state compared to neonatal fibroblasts.
Conclusions:
- The developmental stage of fibroblasts critically influences direct reprogramming efficiency for beta-cell generation.
- Neonatal fibroblasts are a more permissive source for generating beta-cell-like cells.
- Developmental stage-dependent cellular plasticity is key for optimizing reprogramming outcomes.
Related Concept Videos
01:29Somatic to iPS Cell Reprogramming
Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012 for this...
01:22iPS Cell Differentiation
The ability of induced pluripotent stem cells or iPSCs to differentiate into most body cell types has stimulated repair and regenerative medicine research over the past few decades. iPSC-derived blood cells, hepatocytes, beta islet cells, cardiomyocytes, neurons, and other cell types can repair injuries or regenerate damaged tissue in diseases such as diabetes and neurodegenerative disorders.
01:28Forced Transdifferentiation
Transdifferentiation, also known as lineage reprogramming, was first discovered by Selman and Kafatos in 1974 in silkmoths. They observed that the moths’ cuticle-producing cells transformed into salt-producing cells. Many such cases of natural transdifferentiation occur in organisms. In humans, pancreatic alpha cells can become beta cells. In newts, the loss of the eye’s lens causes the pigmented epithelial cells to transdifferentiate into the lens cells.
Artificial transdifferentiation occurs...
Artificial transdifferentiation occurs...
01:23Tissue Renewal without Stem Cells
After cellular or tissue damage, the resident stem cells present in the human body can locally repair and regenerate the damaged tissue or organ. However, even though some tissues do not have stem cells, they can repair and regenerate with the help of pre-existing cells. For example, beta cells of the pancreas and hepatocytes of the liver can divide to renew and regenerate the tissue. Here, both cell division and cell death are well regulated by homeostasis.
However, failure of such a system...
However, failure of such a system...
01:13Induced Pluripotent Stem Cells
Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore called induced pluripotent stem...