INVESTIGADORES
SIMON Maria Victoria
congresos y reuniones científicas
Título:
Proliferation of Retina Neuroblasts in Cocultures of Muller Cells and retinal neurons: Effects of docosahexaenoic acid on photoreceptors differentiation
Autor/es:
POLITI, LUIS; SIMÓN M. VICTORIA; DE LOS SANTOS, BEATRIZ; ROTSTEIN, NORA
Lugar:
Fort Lauderdale
Reunión:
Congreso; ARVO 2009 Annual Meeting; 2009
Institución organizadora:
ARVO
Resumen:
Purpose: Photoreceptor (PHR) loss is the main cause of blindness in
neurodegenerative diseases of the retina. Muller stem cells might be used
as a source of new PHRs as a promising treatment for these diseases. This
requires establishing the conditions by which these cells promote
proliferation of neuroblasts (NB) and their further differentiation into
PHRs.Photoreceptor (PHR) loss is the main cause of blindness in
neurodegenerative diseases of the retina. Muller stem cells might be used
as a source of new PHRs as a promising treatment for these diseases. This
requires establishing the conditions by which these cells promote
proliferation of neuroblasts (NB) and their further differentiation into
PHRs.
Methods: to investigate these questions we used pure primary cultures of
Muller cells or retinal neurons, and neuro-glia co-cultures grown in serum
containing media or in a serum-free medium, supplemented or not with
docosahexaenoic acid (DHA), a lipid molecule that promotes PHR
survival and differentiation. At different developmental times, we
harvested co-culture cells and reseeded them to obtain secondary
subcultures. We then measured BrdU uptake, to determine proliferation,
TUNEL labeling, to quantitate apoptosis and the presence of nestin, NeuN
and Crx and opsin, as stem cell, neuronal and photoreceptor markers,
respectively.to investigate these questions we used pure primary cultures of
Muller cells or retinal neurons, and neuro-glia co-cultures grown in serum
containing media or in a serum-free medium, supplemented or not with
docosahexaenoic acid (DHA), a lipid molecule that promotes PHR
survival and differentiation. At different developmental times, we
harvested co-culture cells and reseeded them to obtain secondary
subcultures. We then measured BrdU uptake, to determine proliferation,
TUNEL labeling, to quantitate apoptosis and the presence of nestin, NeuN
and Crx and opsin, as stem cell, neuronal and photoreceptor markers,
respectively.
Results: NB were absent in primary or secondary pure glial cultures. In
pure neuronal cultures, a significant amount of NB was initially present,
which rapidly differentiated into PHRs and disappeared from the cultures.
In contrast, in neuro-glia co-cultures, NB could be maintained up to 15
days. In secondary cultures obtained from the co-cultures, NB proliferated
and expressed nestin. Moreover, some cells that took up BrdU, later went
on to express NeuN, Crx and opsin, implying that under these conditions
proliferating NB could give rise to new PHRs. Noteworthy, when we
incubated secondary co-cultures with DHA, the percentage of PHR-like
cells expressing opsin and Crx significantly increased with a parallel
reduction in their apoptosis.
cells expressing opsin and Crx significantly increased with a parallel
reduction in their apoptosis.
Conclusions: the interaction of retinal neurons with Muller cells is critical
for the generation and preservation of NB. DHA promoted these NB to
survive and advanced their differentiation into PHRs. Our findings might
provide important tools for successfully using stem cells in the treatment
of retinal diseases.the interaction of retinal neurons with Muller cells is critical
for the generation and preservation of NB. DHA promoted these NB to
survive and advanced their differentiation into PHRs. Our findings might
provide important tools for successfully using stem cells in the treatment
of retinal diseases.NB were absent in primary or secondary pure glial cultures. In
pure neuronal cultures, a significant amount of NB was initially present,
which rapidly differentiated into PHRs and disappeared from the cultures.
In contrast, in neuro-glia co-cultures, NB could be maintained up to 15
days. In secondary cultures obtained from the co-cultures, NB proliferated
and expressed nestin. Moreover, some cells that took up BrdU, later went
on to express NeuN, Crx and opsin, implying that under these conditions
proliferating NB could give rise to new PHRs. Noteworthy, when we
incubated secondary co-cultures with DHA, the percentage of PHR-like
cells expressing opsin and Crx significantly increased with a parallel
reduction in their apoptosis.
cells expressing opsin and Crx significantly increased with a parallel
reduction in their apoptosis.
Conclusions: the interaction of retinal neurons with Muller cells is critical
for the generation and preservation of NB. DHA promoted these NB to
survive and advanced their differentiation into PHRs. Our findings might
provide important tools for successfully using stem cells in the treatment
of retinal diseases.the interaction of retinal neurons with Muller cells is critical
for the generation and preservation of NB. DHA promoted these NB to
survive and advanced their differentiation into PHRs. Our findings might
provide important tools for successfully using stem cells in the treatment
of retinal diseases.