Product Introduction

novastem-MSC Serum-Free Medium For Mesenchymal Stem Cell

• It can be used for the primary cell isolation and subculture of mesenchymal stem cells (MSCs) from multiple sources, such as umbilical cord (UCMSCs), adipose tissue (ADSCs), bone marrow (BMSCs), amniotic membrane (AMSCs), hair follicles (HFSCs), dental pulp (DPSCs), etc., while maintaining the cells' pluripotent differentiation potential.

• It is serum-free, free of any animal-derived components, and antibiotic-free，with stable performance and minimal batch-to-batch variation.
• It features a high cell expansion rate, with the cell count at the same passage being significantly higher than that of similar products on the market.
• It has an independent R&D and production system, ensuring stable supply and high cost-effectiveness.
• It supports clinical-grade / pharmaceutical-grade cell culture.

Usage Instructions

It is recommended that the complete medium be prepared right before use and used up within one month.

If the culture system is small, it is advisable to aliquot the supplement into smaller portions for frozen storage based on actual usage. Prepare the complete medium in proportion when needed to avoid repeated freeze-thaw cycles.

Seeding Density

Cell Passage Time

It is generally around 3 days (72 hours). The proliferation rate of different human Mesenchymal Stem Cells (hMSCs) varies. It is recommended to determine the exact passage time based on cell confluency;

passage is advised when cell confluency reaches approximately 80–90%. Excessive cell confluency (>95%) will affect subsequent cell growth.

Cell Morphology

The cells are spindle-shaped and exhibit a swirling (or vortex-like) growth pattern.

Culture of Mesenchymal Stem Cells

Primary Culture

 Isolate the Wharton's jelly tissue from the umbilical cord and perform primary culture using the tissue block adherence method. Inoculate 0.5g of Wharton's jelly into each T75 culture flask. After culturing in a 37°C cell incubator for 1-2 hours, add 6-8ml of cell culture medium. Supplement with 5ml of medium on days 2-3. Change the medium on days 7-10 by discarding the supernatant and adding 10-15ml of fresh medium. Observe the cell expansion on days 12-14 and proceed with digestion and passage.

 The earliest time for cells to migrate out of the tissue blocks is 5–7 days, and a small number of scattered cells can be observed under the microscope.

 The tissue block adherence efficiency is high, with a large number of tissue blocks adhering to the culture surface.

 A greater number of cells can be harvested.

Subculture

 Compared with similar products on the market, it has higher expansion efficiency. The cells at high passages age more slowly and can support culture for more than 20 passages.

Culture Performance

• The average expansion fold of the Novastem-MSC system from P1 to P5 is 20.7.
• Counted on average from 10 umbilical cords: for example, if approximately 10g of Wharton's jelly tissue is isolated, 2.0×10⁷ cells can be obtained at Passage 0 (P0), and theoretically 1.71×10¹¹ cells can be obtained at Passage 3 (P3).
• When prepared into products at the conventional dosage of 5×10⁷ cells per unit, it can yield 3,420 units in total.

Summary

Cell Passage Time

It is generally around 3 days (72 hours). Different hMSCs vary in their growth rates. It is recommended to determine the exact passage timing based on cell confluency; passage is advised when cell confluency reaches approximately 80%-90%. Excessively high cell confluency (>95%) will affect subsequent cell growth.
The medium volume is 0.2 mL/cm². No coating is required, and no medium change is needed for 3 consecutive days of culture.
When switching from other culture systems to novastem-MSC, the initial cell expansion fold may be relatively low. It is recommended to resuscitate and seed cells with a 1:1 mixture of the original medium and novastem-MSC. After 1 generation of culture, the system can be completely switched to novastem-MSC.

Cell Digestion

It is recommended to use a relatively gentle trypsin, such as Gibco TrypLE™ Express Enzyme (recombinant, 1X). Dilute it with PBS at a 1:1 ratio to prepare a working solution of trypsin before use. The digestion time is 3–5 minutes. After 1–2 minutes of digestion, gently tap the culture vessel; when most cells are observed to detach, terminate the digestion with medium.

Cell Cryopreservation

It is recommended that the final concentration of DMSO be 5%. Before cell digestion, you can first prepare a cryopreservation solution with a volume ratio of DMSO to complete medium of 1:9, and store it pre-cooled at 2–8°C. After the cells to be cryopreserved are mixed with complete medium, slowly add an equal volume of the pre-prepared cryopreservation solution, mix well, and then aliquot for cryopreservation.

Quality control analysis

Product Certificate of Analysis

After passing the release inspection, a corresponding Certificate of Analysis (COA) shall be issued. The main test items include characteristics, physical and chemical properties, safety indicators, and functional indicators.

Detection of Cell Surface Markers

Detection of Surface Markers in Cultured hMSC

• The positive rates of CD73, CD90, CD105, and HLA-ABC are >95.0%
• The positive rates of CD14, CD19, CD34, CD45, and HLA-DRDPDQ are ≤2.0%

Cellular Multilineage Differentiation Potential

Detection of Cellular Immunomodulatory Capacity

The cultured human Mesenchymal Stem Cells (hMSCs) exhibit the following immunomodulatory capabilities:

● Inhibiting lymphocyte proliferation;
● Promoting the upregulation of Regulatory T cells (Treg);
● Promoting the downregulation of T helper cell type 1 (Th1) and T helper cell type 17 (Th17);
● Inhibiting the expression of Tumor Necrosis Factor-α (TNF-α).

Detection of Cell Migration Ability   
Detection of Angiogenic Ability

Karyotype Analysis of Cell Chromosomes

In Vitro Tumorigenicity Assay of Cells

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