Limits...
IGF-1 enhances cell proliferation and survival during early differentiation of mesenchymal stem cells to neural progenitor-like cells.

Huat TJ, Khan AA, Pati S, Mustafa Z, Abdullah JM, Jaafar H - BMC Neurosci (2014)

Bottom Line: To unravel the roles and effects of different growth factors in the differentiation of MSCs into neural lineages, we have differentiated MSCs into neural lineages using different combinations of growth factors.The NPCs derived from IGF-1 treatment also resulted in a better yield after the terminal differentiation into neurons and glial cells than that of the other treated groups.Our results suggested that IGF-1 has a crucial role in the differentiation of MSCs into neuronal lineage by enhancing the proliferation and reducing the apoptosis in the NPCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, School of Medical Sciences, Universiti Sains Malaysia, Jalan Hospital Universiti Sains Malaysia, 16150 Kubang Kerian, Kota Bharu, Kelantan, Malaysia. teejonghuat@gmail.com.

ABSTRACT

Background: There has been increasing interest recently in the plasticity of mesenchymal stem cells (MSCs) and their potential to differentiate into neural lineages. To unravel the roles and effects of different growth factors in the differentiation of MSCs into neural lineages, we have differentiated MSCs into neural lineages using different combinations of growth factors. Based on previous studies of the roles of insulin-like growth factor 1 (IGF-1) in neural stem cell isolation in the laboratory, we hypothesized that IGF-1 can enhance proliferation and reduce apoptosis in neural progenitor-like cells (NPCs) during differentiation of MSCs into NCPs.We induced MSCs differentiation under four different combinations of growth factors: (A) EGF + bFGF, (B) EGF + bFGF + IGF-1, (C) EGF + bFGF + LIF, (D) EGF + bFGF + BDNF, and (E) without growth factors, as a negative control. The neurospheres formed were characterized by immunofluorescence staining against nestin, and the expression was measured by flow cytometry. Cell proliferation and apoptosis were also studied by MTS and Annexin V assay, respectively, at three different time intervals (24 hr, 3 days, and 5 days). The neurospheres formed in the four groups were then terminally differentiated into neuron and glial cells.

Results: The four derived NPCs showed a significantly higher expression of nestin than was shown by the negative control. Among the groups treated with growth factors, NPCs treated with IGF-1 showed the highest expression of nestin. Furthermore, NPCs derived using IGF-1 exhibited the highest cell proliferation and cell survival among the treated groups. The NPCs derived from IGF-1 treatment also resulted in a better yield after the terminal differentiation into neurons and glial cells than that of the other treated groups.

Conclusions: Our results suggested that IGF-1 has a crucial role in the differentiation of MSCs into neuronal lineage by enhancing the proliferation and reducing the apoptosis in the NPCs. This information will be beneficial in the long run for improving both cell-based and cell-free therapy for neurodegenerative diseases.

Show MeSH

Related in: MedlinePlus

Phenotypic analysis of bone marrow mesenchymal stem cells. (A) Primary rat bone marrow MSCs attached to the surface of the flask exhibiting elongation (arrow). (B) Homogenous rat BM MSCs at passage 4 displaying a fibroblast-like appearance and spindle-like arrangement. (C-D) Immunofluorescence of BM MSCs showing a strong expression of CD90 (C) and CD44 (D). (E) BM MSCs showing weak expression of nestin. (F) Immunophenotypes of BM MSCs analysed by flow cytometry. Images were viewed at 400× magnification using an inverted microscope and a fluorescence microscope equipped with a pseudo-fluorescence filter.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4117972&req=5

Fig1: Phenotypic analysis of bone marrow mesenchymal stem cells. (A) Primary rat bone marrow MSCs attached to the surface of the flask exhibiting elongation (arrow). (B) Homogenous rat BM MSCs at passage 4 displaying a fibroblast-like appearance and spindle-like arrangement. (C-D) Immunofluorescence of BM MSCs showing a strong expression of CD90 (C) and CD44 (D). (E) BM MSCs showing weak expression of nestin. (F) Immunophenotypes of BM MSCs analysed by flow cytometry. Images were viewed at 400× magnification using an inverted microscope and a fluorescence microscope equipped with a pseudo-fluorescence filter.

Mentions: BM MSCs have fibroblast-like morphology (Figure 1A) and form whirlpool-like structures when grown to 80% confluence (Figure 1B). Immunofluorescence staining showed bright staining in all MSCs with antibodies against CD90 (Figure 1C) and CD44 (Figure 1D). Only 5.15 ± 0.68% (n = 6) of cells expressed nestin (Figure 1E). No staining was observed against markers of neuronal cell types such as Sox-2, GFAP, and FluoroPan (data not shown). Flow cytometry analysis further showed that BM MSCs were positive for CD90 and CD44 (Figure 1F). BM MSCs were subcultured until passage 4 and showed consistent morphology and phenotype. The phenotypic analyses of the cells isolated are similar to those described in Hong et al. [9], indicating that the BM MSCs isolated for our experiment met the standard criteria of mesenchymal stem cells.Figure 1


IGF-1 enhances cell proliferation and survival during early differentiation of mesenchymal stem cells to neural progenitor-like cells.

Huat TJ, Khan AA, Pati S, Mustafa Z, Abdullah JM, Jaafar H - BMC Neurosci (2014)

Phenotypic analysis of bone marrow mesenchymal stem cells. (A) Primary rat bone marrow MSCs attached to the surface of the flask exhibiting elongation (arrow). (B) Homogenous rat BM MSCs at passage 4 displaying a fibroblast-like appearance and spindle-like arrangement. (C-D) Immunofluorescence of BM MSCs showing a strong expression of CD90 (C) and CD44 (D). (E) BM MSCs showing weak expression of nestin. (F) Immunophenotypes of BM MSCs analysed by flow cytometry. Images were viewed at 400× magnification using an inverted microscope and a fluorescence microscope equipped with a pseudo-fluorescence filter.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4117972&req=5

Fig1: Phenotypic analysis of bone marrow mesenchymal stem cells. (A) Primary rat bone marrow MSCs attached to the surface of the flask exhibiting elongation (arrow). (B) Homogenous rat BM MSCs at passage 4 displaying a fibroblast-like appearance and spindle-like arrangement. (C-D) Immunofluorescence of BM MSCs showing a strong expression of CD90 (C) and CD44 (D). (E) BM MSCs showing weak expression of nestin. (F) Immunophenotypes of BM MSCs analysed by flow cytometry. Images were viewed at 400× magnification using an inverted microscope and a fluorescence microscope equipped with a pseudo-fluorescence filter.
Mentions: BM MSCs have fibroblast-like morphology (Figure 1A) and form whirlpool-like structures when grown to 80% confluence (Figure 1B). Immunofluorescence staining showed bright staining in all MSCs with antibodies against CD90 (Figure 1C) and CD44 (Figure 1D). Only 5.15 ± 0.68% (n = 6) of cells expressed nestin (Figure 1E). No staining was observed against markers of neuronal cell types such as Sox-2, GFAP, and FluoroPan (data not shown). Flow cytometry analysis further showed that BM MSCs were positive for CD90 and CD44 (Figure 1F). BM MSCs were subcultured until passage 4 and showed consistent morphology and phenotype. The phenotypic analyses of the cells isolated are similar to those described in Hong et al. [9], indicating that the BM MSCs isolated for our experiment met the standard criteria of mesenchymal stem cells.Figure 1

Bottom Line: To unravel the roles and effects of different growth factors in the differentiation of MSCs into neural lineages, we have differentiated MSCs into neural lineages using different combinations of growth factors.The NPCs derived from IGF-1 treatment also resulted in a better yield after the terminal differentiation into neurons and glial cells than that of the other treated groups.Our results suggested that IGF-1 has a crucial role in the differentiation of MSCs into neuronal lineage by enhancing the proliferation and reducing the apoptosis in the NPCs.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, School of Medical Sciences, Universiti Sains Malaysia, Jalan Hospital Universiti Sains Malaysia, 16150 Kubang Kerian, Kota Bharu, Kelantan, Malaysia. teejonghuat@gmail.com.

ABSTRACT

Background: There has been increasing interest recently in the plasticity of mesenchymal stem cells (MSCs) and their potential to differentiate into neural lineages. To unravel the roles and effects of different growth factors in the differentiation of MSCs into neural lineages, we have differentiated MSCs into neural lineages using different combinations of growth factors. Based on previous studies of the roles of insulin-like growth factor 1 (IGF-1) in neural stem cell isolation in the laboratory, we hypothesized that IGF-1 can enhance proliferation and reduce apoptosis in neural progenitor-like cells (NPCs) during differentiation of MSCs into NCPs.We induced MSCs differentiation under four different combinations of growth factors: (A) EGF + bFGF, (B) EGF + bFGF + IGF-1, (C) EGF + bFGF + LIF, (D) EGF + bFGF + BDNF, and (E) without growth factors, as a negative control. The neurospheres formed were characterized by immunofluorescence staining against nestin, and the expression was measured by flow cytometry. Cell proliferation and apoptosis were also studied by MTS and Annexin V assay, respectively, at three different time intervals (24 hr, 3 days, and 5 days). The neurospheres formed in the four groups were then terminally differentiated into neuron and glial cells.

Results: The four derived NPCs showed a significantly higher expression of nestin than was shown by the negative control. Among the groups treated with growth factors, NPCs treated with IGF-1 showed the highest expression of nestin. Furthermore, NPCs derived using IGF-1 exhibited the highest cell proliferation and cell survival among the treated groups. The NPCs derived from IGF-1 treatment also resulted in a better yield after the terminal differentiation into neurons and glial cells than that of the other treated groups.

Conclusions: Our results suggested that IGF-1 has a crucial role in the differentiation of MSCs into neuronal lineage by enhancing the proliferation and reducing the apoptosis in the NPCs. This information will be beneficial in the long run for improving both cell-based and cell-free therapy for neurodegenerative diseases.

Show MeSH
Related in: MedlinePlus