Temperature, °C

—|— Cultured and transduced peripheral blood lymphocytes (PBLs) from donor with MPS II —O— Freshly isolated peripheral blood lymphocytes (PBLs) from a normal donor

FIGURE 7.1 Normalized cell volume as a function of temperature for cultured and transduced lymphocytes from a donor with MPS II (■) and freshly isolated mononuclear cells from a normal donor (O). The cells were frozen in IMDM using a cooling rate of 20°C/min. Experimental results are given as an average ±SD for 15 cells (Hubel, A., Darr, T.B., and Norman, J.A., Freezing characteristics of genetically modified lymphocytes for the treatment of MPSII, Cell Transplant, 8, 521, 1999. With Permission).

cells than the fresh cells from a normal donor. In addition, the minimum normalized volume achieved by the cultured cells was significantly less than that typically observed for the fresh cells.

The efflux of water that results in the observed change in cell volume can be analyzed using a nonequilibrium model developed by Mazur.29 Assuming that equilibria of temperature and pressure prevail between the intra- and extracellular media, the following equation can be written:

(V - Vb ) + Vw (vsns ),

1 AHf

' i

1 N

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