A comparison of haematopoietic stem cells from umbilical cord blood and peripheral blood for platelet production in a microfluidic device

被引:11
|
作者
Six, Katrijn R. [1 ,2 ]
Sicot, Geraldine [3 ]
Devloo, Rosalie [1 ]
Feys, Hendrik B. [1 ,2 ]
Baruch, Dominique [3 ,4 ]
Compernolle, Veerle [1 ,2 ,5 ]
机构
[1] Belgian Red Cross Flanders, Transfus Res Ctr, Ottergemsesteenweg 413, B-9000 Ghent, Belgium
[2] Univ Ghent, Fac Med & Hlth Sci, Ghent, Belgium
[3] PlatOD, Paris, France
[4] Univ Paris 05, Sorbonne Paris Cite, INSERM, UMR S1140, Paris, France
[5] Belgian Red Cross Flanders, Blood Serv, Mechelen, Belgium
关键词
haematopoiesis; megakaryopoiesis; platelets; stem cells; tissue engineering; IN-VITRO; HUMAN MEGAKARYOCYTES; CD34(+) CELLS; BONE-MARROW; GENERATION;
D O I
10.1111/vox.12776
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
Background and objectives Several sources of haematopoietic stem cells have been used for static culture of megakaryocytes to produce platelets in vitro. This study compares and characterizes platelets produced in shear flow using precursor cells from either umbilical (UCB) or adult peripheral blood (PB). Materials and methods The efficiency of platelet production of the cultured cells was studied after perfusion in custom-built von Willebrand factor-coated microfluidic flow chambers. Platelet receptor expression and morphology were investigated by flow cytometry and microscopy, respectively. Results Proliferation of stem cells isolated out of UCB was significantly higher (P < 0 center dot 0001) compared to PB. Differentiation of these cells towards megakaryocytes was significantly lower from PB compared to UCB where the fraction of CD42b/CD41 double positive events was 44 +/- 9% versus 76 +/- 11%, respectively (P < 0 center dot 0001). However, in vitro platelet production under hydrodynamic conditions was more efficient with 7 center dot 4 platelet-like particles per input cell from PB compared to 4 center dot 2 from UCB (P = 0 center dot 02). The percentage of events positive for CD42b, CD41 and CD61 was comparable between both stem cell sources. The mean number of receptors per platelet from UCB and PB was similar to that on blood bank platelets with on average 28 000 CD42b, 57 000 CD61 and 5500 CD49b receptors. Microscopy revealed platelets appearing similar to blood bank platelets in morphology, size and actin cytoskeleton, alongside smaller fragments and source megakaryocytes. Conclusion This characterization study suggests that platelets produced in vitro under flow either from UCB or from PB share receptor expression and morphology with donor platelets stored in the blood bank.
引用
收藏
页码:330 / 339
页数:10
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