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Optimizing culture medium composition to improve oligodendrocyte progenitor cell yields in vitro from subventricular zone-derived neural progenitor cell neurospheres.

Franco PG, Pasquini JM, Silvestroff L - PLoS ONE (2015)

Bottom Line: We used Subventricular Zone-derived NSC/NPC primary cultures from newborn mice and compared the effects of different growth factor combinations on cell proliferation and OPC yield.The Platelet Derived Growth Factor-AA and BB homodimers had a positive and significant impact on OPC generation.As a whole, we describe an optimized in vitro method for increasing OPC.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Química Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, and Instituto de Química y Fisicoquímica Biológicas "Profesor Alejandro C. Paladini" (IQUIFIB), UBA-CONICET, Ciudad Autónoma de Buenos Aires, Argentina.

ABSTRACT
Neural Stem and Progenitor Cells (NSC/NPC) are gathering tangible recognition for their uses in cell therapy and cell replacement therapies for human disease, as well as a model system to continue research on overall neural developmental processes in vitro. The Subventricular Zone is one of the largest NSC/NPC niches in the developing mammalian Central Nervous System, and persists through to adulthood. Oligodendrocyte progenitor cell (OPC) enriched cultures are usefull tools for in vitro studies as well as for cell replacement therapies for treating demyelination diseases. We used Subventricular Zone-derived NSC/NPC primary cultures from newborn mice and compared the effects of different growth factor combinations on cell proliferation and OPC yield. The Platelet Derived Growth Factor-AA and BB homodimers had a positive and significant impact on OPC generation. Furthermore, heparin addition to the culture media contributed to further increase overall culture yields. The OPC generated by this protocol were able to mature into Myelin Basic Protein-expressing cells and to interact with neurons in an in vitro co-culture system. As a whole, we describe an optimized in vitro method for increasing OPC.

No MeSH data available.


Related in: MedlinePlus

Heparin effects on cell culture yield from 3 WT and 2 CNP::EGFP mice.Total cell protein concentration (A) and DNA content (B) quantitation in NS obtained from cultures exposed to different growth factor combinations, and in the absence or presence of 1.25 U/L of Heparin in the culture media during proliferation. The error bars in both graphs represent the SD. Data was analyzed with a One-way ANOVA and Dunnet´s post-test, where the “bFGF/EGF” (without Heparin) treatment was established as the control.
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pone.0121774.g005: Heparin effects on cell culture yield from 3 WT and 2 CNP::EGFP mice.Total cell protein concentration (A) and DNA content (B) quantitation in NS obtained from cultures exposed to different growth factor combinations, and in the absence or presence of 1.25 U/L of Heparin in the culture media during proliferation. The error bars in both graphs represent the SD. Data was analyzed with a One-way ANOVA and Dunnet´s post-test, where the “bFGF/EGF” (without Heparin) treatment was established as the control.

Mentions: The NS diameter length was compared between treatments in the absence and presence of 1.25 U/L Heparin, and among WT and Act::EGFP mice strains. There was a tendency to find larger NS in the bFGF/PDGF-BB-treated cultures in the presence of Heparin than in its absence (S5 C Fig.). The comparison of the different growth factor combinations on cell yield in the absence or presence of 1.25 U/L indicated that heparin effectively had an effect on bFGF (Fig. 5A, B and S5 D, E Fig.). Since we were most interested in increasing OPC yields, we analyzed Heparin effects on bFGF/PDGF-BB cultures. Heparin increases the bFGF/PDGF-BB treated culture yield by 40% or 20% when analyzing total proteins and DNA, respectively, and was found to be statistically significant. However, the cell culture yield in the bFGF/PDGF-BB conditions never reached comparable levels to those obtained with the CTL condition (bFGF/EGF).


Optimizing culture medium composition to improve oligodendrocyte progenitor cell yields in vitro from subventricular zone-derived neural progenitor cell neurospheres.

Franco PG, Pasquini JM, Silvestroff L - PLoS ONE (2015)

Heparin effects on cell culture yield from 3 WT and 2 CNP::EGFP mice.Total cell protein concentration (A) and DNA content (B) quantitation in NS obtained from cultures exposed to different growth factor combinations, and in the absence or presence of 1.25 U/L of Heparin in the culture media during proliferation. The error bars in both graphs represent the SD. Data was analyzed with a One-way ANOVA and Dunnet´s post-test, where the “bFGF/EGF” (without Heparin) treatment was established as the control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383518&req=5

pone.0121774.g005: Heparin effects on cell culture yield from 3 WT and 2 CNP::EGFP mice.Total cell protein concentration (A) and DNA content (B) quantitation in NS obtained from cultures exposed to different growth factor combinations, and in the absence or presence of 1.25 U/L of Heparin in the culture media during proliferation. The error bars in both graphs represent the SD. Data was analyzed with a One-way ANOVA and Dunnet´s post-test, where the “bFGF/EGF” (without Heparin) treatment was established as the control.
Mentions: The NS diameter length was compared between treatments in the absence and presence of 1.25 U/L Heparin, and among WT and Act::EGFP mice strains. There was a tendency to find larger NS in the bFGF/PDGF-BB-treated cultures in the presence of Heparin than in its absence (S5 C Fig.). The comparison of the different growth factor combinations on cell yield in the absence or presence of 1.25 U/L indicated that heparin effectively had an effect on bFGF (Fig. 5A, B and S5 D, E Fig.). Since we were most interested in increasing OPC yields, we analyzed Heparin effects on bFGF/PDGF-BB cultures. Heparin increases the bFGF/PDGF-BB treated culture yield by 40% or 20% when analyzing total proteins and DNA, respectively, and was found to be statistically significant. However, the cell culture yield in the bFGF/PDGF-BB conditions never reached comparable levels to those obtained with the CTL condition (bFGF/EGF).

Bottom Line: We used Subventricular Zone-derived NSC/NPC primary cultures from newborn mice and compared the effects of different growth factor combinations on cell proliferation and OPC yield.The Platelet Derived Growth Factor-AA and BB homodimers had a positive and significant impact on OPC generation.As a whole, we describe an optimized in vitro method for increasing OPC.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Química Biológica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, and Instituto de Química y Fisicoquímica Biológicas "Profesor Alejandro C. Paladini" (IQUIFIB), UBA-CONICET, Ciudad Autónoma de Buenos Aires, Argentina.

ABSTRACT
Neural Stem and Progenitor Cells (NSC/NPC) are gathering tangible recognition for their uses in cell therapy and cell replacement therapies for human disease, as well as a model system to continue research on overall neural developmental processes in vitro. The Subventricular Zone is one of the largest NSC/NPC niches in the developing mammalian Central Nervous System, and persists through to adulthood. Oligodendrocyte progenitor cell (OPC) enriched cultures are usefull tools for in vitro studies as well as for cell replacement therapies for treating demyelination diseases. We used Subventricular Zone-derived NSC/NPC primary cultures from newborn mice and compared the effects of different growth factor combinations on cell proliferation and OPC yield. The Platelet Derived Growth Factor-AA and BB homodimers had a positive and significant impact on OPC generation. Furthermore, heparin addition to the culture media contributed to further increase overall culture yields. The OPC generated by this protocol were able to mature into Myelin Basic Protein-expressing cells and to interact with neurons in an in vitro co-culture system. As a whole, we describe an optimized in vitro method for increasing OPC.

No MeSH data available.


Related in: MedlinePlus