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01.09.2009 | Experimental Research | Ausgabe 9/2009

Acta Neurochirurgica 9/2009

Denuded human amniotic membrane seeding bone marrow stromal cells as an effective composite matrix stimulates axonal outgrowth of rat neural cortical cells in vitro

Acta Neurochirurgica > Ausgabe 9/2009
Hong sheng Liang, Peng Liang, Ye Xu, Jia ning Wu, Tao Liang, Xiao ping Xu, En zhong Liu
Wichtige Hinweise


This manuscript is concerning about the efficacy of denuded human amniotic membrane as a matrix for bone marrow stromal cells, and demonstrated that amniotic membrane is a good carrier for bone marrow stromal cell growth and also support rat cortical neuron axonal outgrowth in vitro. The study was carefully designed and executed, and overall conclusions are convincing.
Yuji Shirakata
This paper shows that cortical neurons can extend neutites in vitro on a matrix of denuded human amniotic membrane, and that bone marrow stromal cells seem to enhance the outgrowth. This is relevant to studies of BMSCs and transplantation. It is not clear why DhAM would be needed as a substrate in light of the many other alternatives, so the importance is questionable. However, this matrix is being used for other surgical procedures, and so perhaps this information will be useful.
Michael Beattie
San Francisco, USA



Previous studies have shown that axonal outgrowth in the damaged central nervous system is closely related to the local microenvironment. Transplantation of bone marrow stromal cells (BMSC) or BMSC with some biomaterials has been used to treat various central nervous system diseases with some success. In the current study, we investigated if BMSC on denuded human amniotic membrane (DhAM) as a composite matrix could stimulate axonal outgrowth or not.


After completely removing the cells on the amniotic membrane with a tryptic and mechanical approach, we seeded BMSC on it. The MTS was applied to test the cytotoxicity of DhAM compared with PLGA and PLL. The morphology of the BMSC was observed by light, electronic and laser confocal microscopy. We also used four kinds of substance (PLL, DhAM, BMSC + PLL, BMSC + DhAM) to coculturing with the cortical neurons. Finally, the lengths of axons in each group were studied using the positive axon-specific marker NF–H.


The DhAM was devoid of cellular components and only its intact basement membrane was left. BMSC grew on the substrate and proliferated with a flat to fusiform morphology. In the MTS test, the results indicated that BMSC cultured in DhAM extract had a high survival rate (> 80%). Moreover, the cortical neural axons in the experimental group (BMSC + DhAM) were longer (287.37 ± 12.72 μm) than in the other groups (P < 0.01).


This study demonstrates that the DhAM was a good carrier to support growth of BMSC and BMSC on DhAM was an effective composite matrix to support the outgrowth of the axons of rat cortical neurons in vitro. Future studies of the use of the composite matrix in disorders are planned.

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