Qualifying Osteogenic Potency Assay Metrics for Human Multipotent Stromal Cells: TGF-β2 a Telling Eligible Biomarker
| Autor: | Bianca Mladin, Cornel Balta, Jorge S. Burns, Mariana Ionita, Anca Hermenean, Diana F Becheru, Sami Gharbia, Augustin M Ofiteru, Hildegard Herman |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
Stromal cell
bone marrow Decorin medicine.medical_treatment LS3_12 potency assay Regenerative medicine bone Article Bone and Bones NO Transforming Growth Factor beta2 Osteogenesis TGF-β2 stem cells TGFB2 LS3_7 Humans Medicine multipotent stromal cells RNA Messenger lcsh:QH301-705.5 Cells Cultured osteogensys decorin Extracellular Matrix Proteins business.industry Multipotent Stem Cells adipose tissue gene expression Reproducibility of Results ALPL General Medicine Stem-cell therapy RUNX2 medicine.anatomical_structure lcsh:Biology (General) Cancer research Bone marrow Stromal Cells Stem cell business Biomarkers |
| Zdroj: | Cells, Vol 9, Iss 2559, p 2559 (2020) Cells Volume 9 Issue 12 |
| ISSN: | 2073-4409 |
| Popis: | Potency assays are critical for regenerative medicine, addressing the known challenge of functional heterogeneity among human multipotent stromal cells (hMSC). Necessary laboratory cell expansion allows analysis before implantation in the patient. Levels of induction of five signature gene biomarkers, ALPL, COL1A2, DCN, ELN and RUNX2, constituted a previously reported proof-of-principle osteogenic potency assay. We tested assay modification to enhance reproducibility using six consistent bone marrow derived hBM-MSC and explored applicability to three adipose tissue derived hAT-MSC. Using a potent proprietary osteogenic induction factor, the GUSB/YWAHZ reference gene pair provided real time PCR consistency. The novel assay conditions supported the concept that genes encoding extracellular matrix proteins one week after osteogenic induction were informative. Nonetheless, relatively low induction of COL1A2 and ELN encouraged search for additional biomarkers. TGFB2 mRNA induction, important for osteogenic commitment, was readily quantifiable in both hBM-MSC and hAT-MSC. Combined with DCN, TGFB2 mRNA induction data provided discriminatory power for resolving donor-specific heterogeneity. Histomorphometric decorin and TGF-&beta 2 protein expression patterns in eight-week heterotopic bone implants also discriminated the two non-bone-forming hMSC. We highlight progress towards prompt osteogenic potency assays, needed by current clinical trials to accelerate improved intervention with enhanced stem cell therapy for serious bone fractures. |
| Databáze: | OpenAIRE |
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