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Updated: May 30, 2026
Isolation of Retinal Stem Cells from the Mouse Eye
Published on: September 11, 2010
Chromosomal number aberrations and transformation in adult mouse retinal stem cells in vitro
Meta Djojosubroto1, Frederic Bollotte, Pratyaksha Wirapati
1Unit of Gene Therapy and Stem Cell Biology, Department of Ophthalmology, Jules-Gonin Eye Hospital, Lausanne, Switzerland.
Purpose:
The potential of stem cells (SCs) as a source for cell-based therapy on a wide range of degenerative diseases and damaged tissues such as retinal degeneration has been recognized. Generation of a high number of retinal stem cells (RSCs) in vitro would thus be beneficial for transplantation in the retina. However, as cells in prolonged cultivation may be unstable and thus have a risk of transformation, it is important to assess the stability of these cells.
Methods:
Chromosomal aberrations were analyzed in mouse RSC lines isolated from adult and from postnatal day (PN)1 mouse retinas. Moreover, selected cell lines were tested for anchorage-dependent proliferation, and SCs were transplanted into immunocompromised mice to assess the possibility of transformation.
Results:
Marked aneuploidy occurred in all adult cell lines, albeit to different degrees, and neonatal RSCs were the most stable and displayed a normal karyotype until at least passage 9. Of interest, the level of aneuploidy of adult RSCs did not necessarily correlate with cell transformation. Only the adult RSC lines passaged for longer periods and with a higher dilution ratio underwent transformation. Furthermore, we identified several cell cycle proteins that might support the continuous proliferation and transformation of the cells.
Conclusions:
Adult RSCs rapidly accumulated severe chromosomal aberrations during cultivation, which led to cell transformation in some cell lines. The culture condition plays an important role in supporting the selection and growth of transformed cells.
Insights
Adult retinal stem cells (RSCs) accumulate chromosomal aberrations during cultivation, risking transformation. Neonatal RSCs are more stable, but culture conditions influence adult cell transformation, highlighting the need for careful assessment in cell-based therapies.
Area of Science:
- Stem cell biology
- Regenerative medicine
- Ophthalmology
Background:
- Stem cells (SCs) show promise for treating degenerative diseases and tissue damage, including retinal degeneration.
- Generating numerous retinal stem cells (RSCs) in vitro is crucial for retinal transplantation.
- Prolonged cell cultivation can lead to instability and transformation risk, necessitating stability assessment.
Purpose of the Study:
- To evaluate the chromosomal stability of mouse retinal stem cell (RSC) lines during in vitro cultivation.
- To assess the transformation potential of RSCs under different culture conditions.
- To identify factors influencing RSC stability and transformation.
Main Methods:
- Analyzed chromosomal aberrations in adult and neonatal mouse RSC lines.
- Assessed anchorage-dependent proliferation of selected cell lines.
- Transplanted SCs into immunocompromised mice to monitor for transformation.
Main Results:
- Adult RSCs exhibited significant aneuploidy during cultivation; neonatal RSCs remained stable with normal karyotypes up to passage 9.
- Aneuploidy levels in adult RSCs did not consistently correlate with transformation.
- Transformation occurred in adult RSC lines passaged longer or at higher dilution ratios.
- Identified cell cycle proteins potentially involved in proliferation and transformation.
Conclusions:
- Adult RSCs accumulate severe chromosomal aberrations in vitro, leading to transformation in some cases.
- Culture conditions significantly impact the selection and growth of transformed cells.
- Careful monitoring of RSC stability is essential for safe cell-based therapies.
