Secretion of growth factor in conditioned medium rat bone marrow-derived mesenchymal stem cells (CM-rBMMSC)

• Main Authors: Rinendyaputri, Ratih, Noviantari, Ariyani, Zainuri, Masagus, Nikmah, Uly Alfi, Dany, Frans, Intan, Putri Retno
• Format: Conference Proceeding
• Language: English
• Subjects: Bone marrow; Femur; Growth factors; Proteins; Stem cells; Tibia; Vascular endothelial growth factor
• ISSN: 0094-243X; 1551-7616
• DOI: 10.1063/5.0118570

Mesenchymal stem cells (MSC) has great potential as a therapy for various degenerative diseases. It thought to be able to replace demage cells and secrete various secretome such as growth factors, chemokines, cytokines and extracellular vechicles. Mesenchymal stem cells (MSC) secretome will promote repair mechanism through the paracrine effect. Conditioned medium (CM) containing a complex of MSC-secrete product which can be used as therapy. In this research we are try to characterize the rat bone marror–derived mesechymal stem cells (rBMMSC) protein marker and the growth factor it secretes in CM. In this study we use male wistar rat and isolated MSC from the tibia and femur using the flushing method. Cultures were performed under 5% CO2 (normoxia), MEM culturemedia suplemented with 10% fetal bovine serum (FBS) and characterization of protein marker with CD29+, CD90+ and CD45-using flowcytometry that done passage 1st-4th. Rat BMMSC were starved for 48 hours with basal medium to obtain conditioned medium. Collection of CM-rBMMSC performed by centrifuging 1600 rpm for 5 min filtering by 0,22µl filter unit. Identification of growth factors was carried out by ELISA with protein marker fibroblast growth factor/FGF, nerve growth factor/NGF and vascular endothelial growth factor/VEGF on rBMMSC passage 4th. The result show that protein marker of rBMMSC were (94,68%CD90-APC; 82,010%CD29-FITC; 8,16%CD45-PE),(98,78%CD90-APC; 89,54%CD29-FITC; 8,70%CD45-PE), (99,93%CD90-APC; 99,66%CD29-FITC; 8,42%CD45-PE) and (99,9%CD90-APC; 99,64%CD29-FITC; 2,4%CD45PE) at passage 1st, 2nd,3th and 4th. On the 4th passage of rBMMSC we got 16,83pg/ml NGF, 905,38pg/ml FGF and 0pg/ml VEGF. This study show that CM-rBMMSC can be an alternative cell-free theraphy.