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BMC Biotechnology

, 16:89

Cellular and tissue engineering


BackgroundThe human body has an extensive capacity to regenerate bone tissue after trauma. However large defects such as long bone fractures of the lower limbs cannot be restored without intervention and often lead to nonunion. Therefore, the aim of the present study was to assess the pool and biological functions of human mesenchymal stromal cells hMSCs isolated from different bone marrow locations of the lower limbs and to identify novel strategies to prime the cells prior to their use in bone fracture healing. Following, bone marrow from the ilium, proximal femur, distal femur and proximal tibia was aspirated and the hMSCs isolated. Bone marrow type, volume, number of mononuclear cells-hMSCs and their self-renewal, multilineage potential, extracellular matrix ECM production and surface marker profiling were analyzed. Additionally, the cells were primed to accelerate bone fracture healing either by using acoustic stimulation or varying the initial hMSCs isolation conditions.

ResultsWe found that the more proximal the bone marrow aspiration location, the larger the bone marrow volume was, the higher the content in mononuclear cells-hMSCs and the higher the self-renewal and osteogenic differentiation potential of the isolated hMSCs were. Acoustic stimulation of bone marrow, as well as the isolation of hMSCs in the absence of fetal bovine serum, increased the osteogenic and ECM production potential of the cells, respectively.

ConclusionWe showed that bone marrow properties change with the aspiration location, potentially explaining the differences in bone fracture healing between the tibia and the femur. Furthermore, we showed two new priming methods capable of enhancing bone fracture healing.

KeywordsBone marrow hMSCs Acoustic stimulation Cell priming AbbreviationsALPAlkaline phosphatase

BFHBone fracture healing


BMBone marrow

CFUColony forming unit

CFU-ObColony forming unit osteoblast


ECMExtracellular matrix

FBSFetal bovine serum

GMGrowth media

hMSCsHuman mesenchymal stromal cells

MNCsMononuclear cells

P1Passage 1

P2Passage 2

PDPopulation doubling

SFSerum free

THATotal hip arthroplasty

TKATotal knee arthroplasty

α-MEMα-minimal essential media

Electronic supplementary materialThe online version of this article doi:10.1186-s12896-016-0318-1 contains supplementary material, which is available to authorized users.

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Autor: Corina Adriana Ghebes - Maaike Vera Jasmijn Braham - Adelgunde Veronica Clemens Maria Zeegers - Auke Jan Sijbe Renard


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