125both USP (ch. 1223) and EP (Sect. 5.1.6). At present, there are some solutions for
rapid microbiological testing, but all have advantages and disadvantages. The most
comparable to pharmacopoeial methods are cultivation methods based on CO 2
detection (BACTEC™, Becton Dickinson, and BacT/ALERT ® , bioMérieux). Other
kits using DNA detection tests (e.g., LightCycler ® SeptiFast Test, Roche) may be
more challenging for validation, because they may not detect all possible contami-
nating organisms. Fluorescent cytometry tests (Scan RDI ® , AES Chemunex) pro-
vide ultrarapid detection of microorganisms (90 min), but are very expensive and
typically used by large pharmacological companies.
Mycoplasmas are microorganisms without a cell wall, which can pass through
0.2-μm fi lters used for sterilization. Mycoplasma detection is required for cell cul-
ture according to European, US, and Japanese pharmacopoeias. Although there are
kits to detect mycoplasma based on DNA, standard tests are still used to confi rm
mycoplasma contamination. There are two types of tests to confi rm mycoplasma
contamination. The fi rst is inoculation of cell culture samples on solid agar or in
Fig. 6.6 Flowchart of GMP-compliant production of MSCs for clinical application. All steps
from donor selection to storage and delivery should be controlled and recorded
6 Production of Clinical-Grade Mesenchymal Stem Cells