Limits...
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

Analying the cell yield by measuring different parameters in Actin::EGFP (A-F) and WT (B) mice NS cultures.A) The NS radius length was measured under the different experimental conditions. B) Comparison of EGFP and Höechst fluorescence as a function of total proteins in NS cultures derived from WT and Actin::EGFP mice. C) The radius length, DNA content and EGFP fluorescence are expressed as a function of the protein concentration. D-F) Whole NS extracts were analyzed according to the total cell protein concentration (D), DNA content (E) and EGFP fluorescence (F) for different experimental conditions. Data in A, D-F were analyzed with a One-way ANOVA and Dunnet´s post-test, where the EGF/bFGF treatment was established as the control and error bars represent the SD. * = p < 0.05, ** = p < 0.01, *** = p < 0.001.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4383518&req=5

pone.0121774.g002: Analying the cell yield by measuring different parameters in Actin::EGFP (A-F) and WT (B) mice NS cultures.A) The NS radius length was measured under the different experimental conditions. B) Comparison of EGFP and Höechst fluorescence as a function of total proteins in NS cultures derived from WT and Actin::EGFP mice. C) The radius length, DNA content and EGFP fluorescence are expressed as a function of the protein concentration. D-F) Whole NS extracts were analyzed according to the total cell protein concentration (D), DNA content (E) and EGFP fluorescence (F) for different experimental conditions. Data in A, D-F were analyzed with a One-way ANOVA and Dunnet´s post-test, where the EGF/bFGF treatment was established as the control and error bars represent the SD. * = p < 0.05, ** = p < 0.01, *** = p < 0.001.

Mentions: A multiple analysis of culture yields was performed by first measuring Act::EGFP NS radii after treatment with the different growth factors combinations. Adding bFGF on its own, or together with PDGF generates small sized NS compared to the CTL (Fig. 2A). There was a tendency to find larger NS when PDGF was added to the bFGF condition, being PDGF-BB more effective on increasing the NS size than PDGF-AA.


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)

Analying the cell yield by measuring different parameters in Actin::EGFP (A-F) and WT (B) mice NS cultures.A) The NS radius length was measured under the different experimental conditions. B) Comparison of EGFP and Höechst fluorescence as a function of total proteins in NS cultures derived from WT and Actin::EGFP mice. C) The radius length, DNA content and EGFP fluorescence are expressed as a function of the protein concentration. D-F) Whole NS extracts were analyzed according to the total cell protein concentration (D), DNA content (E) and EGFP fluorescence (F) for different experimental conditions. Data in A, D-F were analyzed with a One-way ANOVA and Dunnet´s post-test, where the EGF/bFGF treatment was established as the control and error bars represent the SD. * = p < 0.05, ** = p < 0.01, *** = p < 0.001.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0121774.g002: Analying the cell yield by measuring different parameters in Actin::EGFP (A-F) and WT (B) mice NS cultures.A) The NS radius length was measured under the different experimental conditions. B) Comparison of EGFP and Höechst fluorescence as a function of total proteins in NS cultures derived from WT and Actin::EGFP mice. C) The radius length, DNA content and EGFP fluorescence are expressed as a function of the protein concentration. D-F) Whole NS extracts were analyzed according to the total cell protein concentration (D), DNA content (E) and EGFP fluorescence (F) for different experimental conditions. Data in A, D-F were analyzed with a One-way ANOVA and Dunnet´s post-test, where the EGF/bFGF treatment was established as the control and error bars represent the SD. * = p < 0.05, ** = p < 0.01, *** = p < 0.001.
Mentions: A multiple analysis of culture yields was performed by first measuring Act::EGFP NS radii after treatment with the different growth factors combinations. Adding bFGF on its own, or together with PDGF generates small sized NS compared to the CTL (Fig. 2A). There was a tendency to find larger NS when PDGF was added to the bFGF condition, being PDGF-BB more effective on increasing the NS size than PDGF-AA.

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