Tags

Type your tag names separated by a space and hit enter

[Research of human keratinocyte growth factor promoting proliferation and differentiation of human neural stem cells].
Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013 Oct; 27(10):1246-51.ZX

Abstract

OBJECTIVE

To study the effects of the human keratinocyte growth factor 2 (hKGF-2) on the survival and differentiation of human neural stem cells (hNSCs).

METHODS

The hNSCs at 17 passages preserved in liquid nitrogen were resuscitated and cultured for 7 days with normal methods to form neural spheres. The specific Nestin antigen and differentiated cells antigen were identified using immunohistochemistry technology. Some concentrated hNSCs were incubated in 12-well culture plate with 1 mL basic medium [(DMEM/F12 + N2 (1 : 100) + epidermal growth factor (EGF) (20 ng/mL)] and divided into 7 groups, 6 wells each group. hKGF-2 (0, 10, 30, 60, 90, and 120 ng/mL) and bFGF (10 ng/mL) were added in groups A (control), B, C, D, E, F, and G, respectively. The neurospheres and the cell number were recorded for analyzing growth and multiplication of neural spheres. Some concentrated hNSCs were incubated in 6-well culture plate (cover glass coated with polylysine) with 3 mL DMEM/F12 medium and divided into 4 groups, 6 wells each group. N2 (1 : 100), N2 (1 : 100) + hKGF-2 (90 ng/mL), FBS (1 : 20), and FBS (1 : 20) + hKGF-2 (90 ng/mL) were added in groups A1, B1, C1, and D1, respectively. Then, the growth and multiplication of neural spheres were observed during culture; the separated neural spheres was identified and analyzed with indirect immunofluorescence and flow cytometry.

RESULTS

Reanimated hNSCs could form neural spheres containing a lot of Nestin antigen; differentiated cells by induction expressed the specific antigens of neurofilament 200 (NF-200) and glial fibrillary acidic protein (GFAP). At 7 days after culture, enlarged neural spheres were observed in each group. The neurospheres and the cell number of hNSCs increased with increased concentration of hKGF-2, showing a gradually increasing tendency; they were significantly higher in groups E, F, and G than that in groups A, B, C, and D (P < 0.05); significant differences were found among groups B, C, and D (P < 0.05), but no significant difference between groups A and B, and among groups E, F, and G (P > 0.05). After induction in vitro, the cell growth showed a progressive increase, significant difference was found among groups (P < 0.05); the percentage of NF-200 positive cells in group B1 was significantly higher than that in the other 3 groups (P < 0.05); the percentage of GFAP positive cells in group B1 was significantly lower than that in the other 3 groups (P < 0.05), but no significant difference among groups A1, C1, and D1 (P > 0.05). At 14 days after culture, cell growth reached the peak, which were mainly astero-cells.

CONCLUSION

The hNSCs are pure after incubated to 17 passages in vitro. hKGF-2 can promote the clone and the growth of differentiated cells, and increase the proportion of neuron.

Authors+Show Affiliations

Department of Orthopaedics, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.Department of Orthopaedics, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.Department of Orthopaedics, First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, PR China.

Pub Type(s)

English Abstract
Journal Article
Research Support, Non-U.S. Gov't

Language

chi

PubMed ID

24397140

Citation

Yang, Yunkang, et al. "[Research of Human Keratinocyte Growth Factor Promoting Proliferation and Differentiation of Human Neural Stem Cells]." Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery, vol. 27, no. 10, 2013, pp. 1246-51.
Yang Y, Jiang D, Yang M. [Research of human keratinocyte growth factor promoting proliferation and differentiation of human neural stem cells]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013;27(10):1246-51.
Yang, Y., Jiang, D., & Yang, M. (2013). [Research of human keratinocyte growth factor promoting proliferation and differentiation of human neural stem cells]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery, 27(10), 1246-51.
Yang Y, Jiang D, Yang M. [Research of Human Keratinocyte Growth Factor Promoting Proliferation and Differentiation of Human Neural Stem Cells]. Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2013;27(10):1246-51. PubMed PMID: 24397140.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - [Research of human keratinocyte growth factor promoting proliferation and differentiation of human neural stem cells]. AU - Yang,Yunkang, AU - Jiang,Dianming, AU - Yang,Mingming, PY - 2014/1/9/entrez PY - 2014/1/9/pubmed PY - 2014/7/6/medline SP - 1246 EP - 51 JF - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery JO - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi VL - 27 IS - 10 N2 - OBJECTIVE: To study the effects of the human keratinocyte growth factor 2 (hKGF-2) on the survival and differentiation of human neural stem cells (hNSCs). METHODS: The hNSCs at 17 passages preserved in liquid nitrogen were resuscitated and cultured for 7 days with normal methods to form neural spheres. The specific Nestin antigen and differentiated cells antigen were identified using immunohistochemistry technology. Some concentrated hNSCs were incubated in 12-well culture plate with 1 mL basic medium [(DMEM/F12 + N2 (1 : 100) + epidermal growth factor (EGF) (20 ng/mL)] and divided into 7 groups, 6 wells each group. hKGF-2 (0, 10, 30, 60, 90, and 120 ng/mL) and bFGF (10 ng/mL) were added in groups A (control), B, C, D, E, F, and G, respectively. The neurospheres and the cell number were recorded for analyzing growth and multiplication of neural spheres. Some concentrated hNSCs were incubated in 6-well culture plate (cover glass coated with polylysine) with 3 mL DMEM/F12 medium and divided into 4 groups, 6 wells each group. N2 (1 : 100), N2 (1 : 100) + hKGF-2 (90 ng/mL), FBS (1 : 20), and FBS (1 : 20) + hKGF-2 (90 ng/mL) were added in groups A1, B1, C1, and D1, respectively. Then, the growth and multiplication of neural spheres were observed during culture; the separated neural spheres was identified and analyzed with indirect immunofluorescence and flow cytometry. RESULTS: Reanimated hNSCs could form neural spheres containing a lot of Nestin antigen; differentiated cells by induction expressed the specific antigens of neurofilament 200 (NF-200) and glial fibrillary acidic protein (GFAP). At 7 days after culture, enlarged neural spheres were observed in each group. The neurospheres and the cell number of hNSCs increased with increased concentration of hKGF-2, showing a gradually increasing tendency; they were significantly higher in groups E, F, and G than that in groups A, B, C, and D (P < 0.05); significant differences were found among groups B, C, and D (P < 0.05), but no significant difference between groups A and B, and among groups E, F, and G (P > 0.05). After induction in vitro, the cell growth showed a progressive increase, significant difference was found among groups (P < 0.05); the percentage of NF-200 positive cells in group B1 was significantly higher than that in the other 3 groups (P < 0.05); the percentage of GFAP positive cells in group B1 was significantly lower than that in the other 3 groups (P < 0.05), but no significant difference among groups A1, C1, and D1 (P > 0.05). At 14 days after culture, cell growth reached the peak, which were mainly astero-cells. CONCLUSION: The hNSCs are pure after incubated to 17 passages in vitro. hKGF-2 can promote the clone and the growth of differentiated cells, and increase the proportion of neuron. SN - 1002-1892 UR - https://www.unboundmedicine.com/medline/citation/24397140/[Research_of_human_keratinocyte_growth_factor_promoting_proliferation_and_differentiation_of_human_neural_stem_cells]_ DB - PRIME DP - Unbound Medicine ER -