Impact of humanised isolation and culture conditions on stemness and osteogenic potential of bone marrow derived mesenchymal stromal cells
| العنوان: | Impact of humanised isolation and culture conditions on stemness and osteogenic potential of bone marrow derived mesenchymal stromal cells |
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| المؤلفون: | Daniela Elena Costea, Pierre Layrolle, Jan E. Brinchmann, Jérôme Amiaud, Kamal Mustafa, Tommy A. Karlsen, Hassan R. W. Ali, Salwa Suliman, Samih Mohamed-Ahmed |
| المصدر: | Scientific Reports, Vol 9, Iss 1, Pp 1-18 (2019) Scientific Reports |
| بيانات النشر: | Nature Publishing Group, 2019. |
| سنة النشر: | 2019 |
| مصطلحات موضوعية: | Stromal cell, Cell Culture Techniques, lcsh:Medicine, Bone Marrow Cells, Cell Separation, Mice, SCID, Mesenchymal Stem Cell Transplantation, Article, 03 medical and health sciences, Mice, 0302 clinical medicine, In vivo, Mice, Inbred NOD, Osteogenesis, medicine, Animals, Humans, lcsh:Science, 030304 developmental biology, 0303 health sciences, Multidisciplinary, Chemistry, Mesenchymal stem cell, lcsh:R, Cell Differentiation, Mesenchymal Stem Cells, Translational research, Molecular biology, In vitro, Stem-cell research, medicine.anatomical_structure, Cell culture, 030220 oncology & carcinogenesis, Heterografts, Platelet lysate, lcsh:Q, Bone marrow, Stem cell |
| الوصف: | Therapeutic potential of human bone marrow stromal/stem cells (hBMSC) must be developed using well defined xenogenic-free conditions. hBMSC were isolated from healthy donors (n = 3) using different isolation and expansion methods. Donor I was isolated and expanded by either bone marrow directly seeded and cells expanded in 10% AB human serum (AB) +5 ng/ml fibroblast growth factor-2 (FGF2) [Direct(AB + FGFlow)] or Ammonium-Chloride-Potassium Lysing Buffer was used before the cells were expanded in 10% AB +5 ng/ml FGF-2 [ACK(AB + FGFlow)] or Lymphoprep density gradient medium was used before the cells were expanded in 10% AB +5 ng/ml FGF2 [Lympho(AB + FGFlow)] or bone marrow directly seeded and cells expanded in 10% pooled platelet lysate plasma (PL) + heparin (2 I/U/mL) [Direct(PL)]. Groups for donors II and III were: Direct(AB + FGFlow) or 10% AB +10 ng/ml FGF2 [Direct(AB + FGFhigh)] or Direct(PL). HBMSCs were assessed for viability, multi-potency, osteogenic, inflammatory response and replicative senescence in vitro after 1 and 3 weeks. Pre-selected culture conditions, Direct(AB + FGFhigh) or Direct(PL), were seeded on biphasic calcium phosphate granules and subcutaneously implanted in NOD/SCID mice. After 1 and 11 weeks, explants were analysed for inflammatory and osteogenic response at gene level and histologically. To identify implanted human cells, in situ hybridisation was performed. hBMSC from all conditions showed in vitro multi-lineage potency. hBMSCs expanded in PL expressed stemness markers in vitro at significantly higher levels. Generally, cells expanded in AB + FGF2 conditions expressed higher osteogenic markers after 1 week both in vitro and in vivo. After 11 weeks in vivo, Direct(AB + FGFhigh) formed mature ectopic bone, compared to immature mineralised tissues formed by Direct(PL) implants. Mouse responses showed a significant upregulation of IL-1α and IL-1β expression in Direct(PL). After 1 week, human cells were observed in both groups and after 11 weeks in Direct(AB + FGFhigh) only. To conclude, results showed a significant effect of the isolation methods and demonstrated a relatively consistent pattern of efficacy from all donors. A tendency of hBMSC expanded in PL to retain a more stem-like phenotype elucidates their delayed differentiation and different inflammatory expressions. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 2045-2322 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5ce9ead175f7df267e50a29e1162eefcTest http://link.springer.com/article/10.1038/s41598-019-52442-9Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....5ce9ead175f7df267e50a29e1162eefc |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 20452322 |
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