9. Stain cells harvested (about 10⁶ cells) for expression of cell

surface markers (PE-conjugated anti-human CD73, FITC-

conjugated anti-human CD90, PE-conjugated anti-human

CD105, FITC-conjugated anti-human CD14, PE-conjugated

anti-human CD19, FITC-conjugated anti-human CD34,

APC-conjugated anti-human CD45, and PE-conjugated anti-

human HLA-DR CD19) for immunophenotype characteriza-

tion by fluorescence-activated cell sorting (FACS) according to

[11] (Fig. 9).

4

Notes

1. Cell viability should be at least 95%. Mix a sample of cell

suspension with Trypan blue solution at 1:1 ratio and count

cells. Cells should be counted within 3 min. Non-viable cells

are stained blue.

2. pH sensor should be fully immersed in calibration solutions

(pH 4.00 and pH 7.01, respectively) for several times.

3. NaOH solution has strong causticity. Please exercise safety

measures at all times.

4. Tube 1 for material feed, tube 2 for complete liquid discharge

and harvest, tube 3 for medium exchange (this tube is fitted

with a filter device to prevent microcarriers from been dis-

charged), tube 4 for sampling, tube 5 for air ring sparger,

tube 6 for direct air vent, tube 7a and 7b for pressure testing

(to be connected with one short silicon tubing after pressure

testing), and tube 8 for air vent condenser.

Fig. 9 Immunophenotypic characterization of hUCMSCs. These cells expressed antigens CD73, CD90, and

CD105, but not antigens CD14, CD-19, HLA-DR, CD34, and CD45

Large-Scale Expansion of UCMSCs with Microcarrier Tablets

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