In vitro differentiation of adult adipose mesenchymal stem cells into retinal progenitor cells.
BACKGROUND
It has been previously shown that adult mesenchymal stem cells (MSCs) differentiate into neural progenitor cells (NPCs) and that the differentiation process was completed in 24-48 h. In this previous study, MSCs from a bone marrow or fat source were co-incubated with homologous autoaggressive cells (ECs) against nerve tissue, and these NPCs were successfully used in human regenerative therapeutic approaches. The present study was conducted to investigate whether a similar differentiation method could be used to obtain autologous retinal progenitor cells (RPCs).


METHODS
Human Th1 cells against retinal tissue were obtained by challenging human blood mononuclear cells with an eye lysate of bovine origin; negative selection was performed using a specific immunomagnetic bead cocktail. Fat MSCs were obtained from a human donor through mechanical and enzymatic dissociation of a surgical sample. The ECs and MSCs were co-cultured in a serum-free medium without the addition of cytokines for 0, 24, 48 and 72 h. The plastic adherent cells were morphologically examined using inverted-phase microscopy and characterized by immunofluorescent staining using antibodies against Pax 6, TUBB3, GFAP, Bestrophin 2, RPE 65, OPN1 SW, and rhodopsin antigens.


RESULTS
The early signs of MSC differentiation into RPCs were observed at 24 h of co-culture, and the early differentiated retinal linage cells appeared at 72 h (neurons, rods, Müller cells, retinal ganglion cells and retinal pigmented epithelial cells). These changes increased during further culture.


CONCLUSIONS
The results reported here support the development of a method to obtain a large number of autologous adult RPCs, which could be used to treat different retinopathies.
Center for Research in Tissue Engineering and Cellular Therapies, Maimónides University, Buenos Aires, Argentina.
Gustavo A
GA
Moviglia
Nahuel
N
Blasetti
Jorge O
JO
Zarate
David E
DE
Pelayes
D000273:Adipose Tissue / Q000166:cytology* / Q000378:metabolism; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D002469:Cell Separation; D018920:Coculture Techniques; D016895:Culture Media, Serum-Free; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D008858:Microscopy, Phase-Contrast; D012160:Retina / Q000166:cytology* / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D018417:Th1 Cells / Q000166:cytology; D014019:Tissue Donors
D015415:Biomarkers; D016895:Culture Media, Serum-Free