Serum-free, xeno-free and animal component-free systems for the culture and differentiation of human mesenchymal stem cells Biological Industries

What works With MSC Attachment Solution With Corning CellBind® Dissociation Recombinant Trypsin Solutions +/- EDTA With Platelet Lysate With ACF recombinant laminin Dissociation Recombinant Trypsin Solution +/- EDTA MSCgo™ With Adipogenic Differentiation Medium MSCgo™ Chondrogenic Differentiation Medium MSCgo™ Osteogenic Differentiation Medium MSCgo™ Rapid Osteogenic Differenti

Seamless transition from research to the clinic Human mesenchymal stem cells (hMSC) are multipotent adult stem cells present in a variety of tissue niches in the human body. hMSC have advantages over other stem cell types due to the broad variety of their tissue sources, for being immunoprivileged, and for their ability to specifically migrate to tumors and wounds in vivo. Due to these traits hMSC have become desirable tools in tissue engineering and cell therapy. In most clinical applications hMSC are expanded in vitro before use. The quality of the culture medium and its performance are particularly...

Product Name MSC NutriStem® XF Xeno-Free, Serum-Free Culture System Size Storage Phenol Red-Free Basal Medium MSC Attachment Solution 05-752-1F 1ml 2-8°C Xeno-free, serum-free culture system, specially designed to support the growth of hMSC from various sources. Advantages Excellent performance —> Superior isolation and cell growth —> Superior maintenance of hMSC characteristics Suitable for research and clinical applications —> Produced under cGMP conditions —> FDA drug master file available —> Used in clinical trials worldwide Defined, serum-free, xeno-free medium —> Reproducible and...

hMSC from various sources (hMSC-PL, hMSC-AT, hMSC-WJ, hMSC-BM) can be efficiently isolated using MSC NutriStem® XF on pre-coated dishes. Addition of 2-2.5% human AB serum may be required for certain tissues. Using MSC NutriStem® XF for isolation of hMSC enhances purity of MSC populations in earlier passages and increases the number of hMSC in comparison to FBS-containing medium. with human AB serum without serum MSC NutriStem® XF Serum-containg medium Figure 1: hMSC were isolated from frozen crude placenta under SF, XF culture conditions (MSC XF on pre-coated plates with MSC Attachment Solution,...

Viable isolated cells (x106) Serum-containing medium MSC NutriStem® XF +2% human AB serum Weiss medium (composed of 2% FBS) Day 2 post initial isolation Representative images MSC NutriStem® XF (supplemented with 2% human AB serum) Day 7 post initial isolation Total cell count Live Dead 99.2% Weiss medium (composed of 2% FBS) Serum-containing medium Figure 4: hMSC were initially isolated from 4 independent human umbilical cords utilizing MSC NutriStem® XF supplemented with 2% human AB serum on pre-coated plates with MSC Attachment Solution in comparison to serum-containing medium. A. Comparing...

Superior proliferation of hMSC hMSC cultured in MSC NutriStem® XF exhibit higher proliferation rate and long term growth in comparison to competitors’ media. Serum-containing medium Commercial serum-containing medium Figure 8: Expansion of hMSC-AT in MSC NutriStem® XF and commercially available XF, SF, and serum-containing media. Cells were cultured in plates, pre-coated with MSC Attachment Solution. Initial seeding was 5000 cells/cm2 for each of the tested media (day 0). Cells were counted at day 3 in each passage. Days post seeding Figure 6: hMSC-BM were cultured in MSC NutriStem® XF in comparison...

Cell morphology Self-renewal potential Typical fibroblast-like cells morphology was obtained when using MSC NutriStem® XF. hMSC cultured in MSC NutriStem® XF maintain their selfrenewal potential. normal morphology Figure 13: hMSC-BM and AT expanded in MSC NutriStem® XF for 3-5 passages prior to 14 day CFU-F assay. Representative images of colonies stained with 0.5% crystal violet (x100). abnormal morphology Figure 11: Expansion of hMSC-AT in MSC NutriStem® XF, competitor XF medium and competitor SF medium (day 0). Initial seeding was 5000 cells/cm2 for each of the tested media. Images (x200)...

Differentiation potential Surface markers profile hMSC cultured in MSC NutriStem® XF maintain their trilineage differentiation potential. hMSC expanded in MSC NutriStem® XF kept their classical profile of MSC markers; stained for MSC positive surface markers and did not stain for hematopoietic markers. hMSC-BM hMSC-PL Chondrocyte Alcian blue 92.9% Figure 16: immunophenotype results of hMSC-PL after culturing in each medium. Purer hMSC population is achieved using MSC Chondrocyte Alcian blue Figure 15: hMSC-BM and hMSC-AT were expanded in MSC NutriStem® XF for 3-5 passages prior to differentiation....

Karyotyping Normal karyotypes of hMSC-BM (46, XY) hMSC-AT (46,XX) and hMSC-CT (46, XX) were observed after long term culturing in MSC NutriStem® XF. MSC NutriStem® XF Complete Medium Stability The complete MSC NutriStem® XF is stable for 30 days at 2- 8°C. No significant differences of hMSC proliferation were observed between fresh and 30 days old complete medium. P6 PD18 46, XX Figure 18: Growth curve of hMSC-BM cultured in MSC NutriStem® XF. Results were taken from 2 batches: batch No. 1: 7.5 months from production and batch 4A: 2.5 months from production. A complete medium was prepared 30...

Key References • C. Elseberg et al. The Challenge of Human Mesenchymal Stromal Cell Expansion: Current and Prospective Answers. New Insights into Cell Culture Technology, Dr. Sivakumar Joghi Thatha Gowder (Ed.), InTech. 2017 • S. Bobis-Wozowicz et al. Diverse impact of xeno-free conditions on biological and regenerative properties of hUC-MSCs and their extracellular vesicles. Journal of Molecular Medicine, 2016 • K.Y. Tan et al. Serum-free media formulations are cell line–specific and require optimization for microcarrier culture. Cytotherapy, 2015 • M.Pokrywczynska et al., Transdifferentiation...