Direct induction versus neurosphere induction of neural cells from rat adipose stromal cells: a comparative study
10.3760/cma.j.issn.1671-8925.2009.08.009
- VernacularTitle:不同方法诱导大鼠脂肪基质细胞定向分化为神经细胞的比较研究
- Author:
Yan-Ping TANG
1
;
Xiao-Dan JIANG
;
Hong-Tian ZHANG
;
Yu-Xi ZOU
;
Ru-Xiang XU
Author Information
1. 南方医科大学珠江医院
- Keywords:
Adipose-derinved stromal cells;
Cell induction;
Cell differentiation;
Neuron-like cells
- From:
Chinese Journal of Neuromedicine
2009;8(8):790-793
- CountryChina
- Language:Chinese
-
Abstract:
Objective To enhance the differentiation efficiency of adipose-derived stromal cells (ADSCs) into neural cells. Methods ADSCs were isolated from the adipose tissue of SD rats and induced to differentiate into neural cells using two different protocols, namely direct induction (group A) and neurosphere induction (group B) protocols. For direct induction, the ADSCs were cultured in Neurobasal medium containing fetal bovine serum (FBS) and neurotrophic factors (NTs). Neurosphere induction protocol was carried out by culturing the ADSCs in serum-free Neurobasal medium to induce the formation of neurospheres, which were further induced in serum- and NT-containing Nenrobasal medium into neural cells. Morphological observation and immunohistochemistry for nestin, β-tubulin Ⅲ, MAP2ab and GFAP were used to identify the differentiated cells. Results Primary cultured ADSCs appeared polymorphous, from which the mixed cells were removed by adherent culture. The fifth passage of ADSCs showed homogenous morphology in regular alignment, and the phenotype could be maintained after further passaging. Immunohistochemistry showed that the two induction protocols both resulted in high expression of nestin and β-tubulin Ⅲ in the induced ADSCs, and nestin expression in group A reached the peak level of(73.8±6.5)% at 6 h of the induction, followed by rapid reduction to (50.3±3.8)% at 24 h and (10.5±30)% at 72 h, becoming almost undetectable afterwards;β-tubulin Ⅲ expression occurred at 24 h following the induction [(33.5±6.6)%] and reached the peak level of (84.3±33)% at 1 week. In group B, high nestin expression persisted throughout the neurosphere stage, while low levels of β-tubulin Ⅲ expression was detected during the neurosphere stage [(14.1±3.3)% at 7 days after neurosphere formation], but its expression increased obviously with time after neurosphere differentiation [(46.4±6.1)% at 3 days after the differentiation. In group B, 24.5% of the cells were found positive for MAP2ab after the differentiation of the neurospheres into neural cells. Conclusion ADSCs can be induced into neural cells under appropriate conditions. Both of the induction protocols can obtain high rate of nestin- and β-tubulin Ⅲ -positive cells, and the neurosphere induction protocol produces more stable nestin expression in the differentiated cells, which contain a proportion of MAP2ab-positive cells.
