PMID- 2379413
OWN - NLM
STAT- MEDLINE
DCOM- 19900911
LR  - 20190912
IS  - 0011-2240 (Print)
IS  - 0011-2240 (Linking)
VI  - 27
IP  - 3
DP  - 1990 Jun
TI  - The cryobiology of rat and human dendritic cells: preservation and destruction of 
      membrane integrity by freezing.
PG  - 269-78
AB  - Dendritic cells (DCs) are now regarded as specialized leucocytes with distinctive 
      morphological and functional characteristics as accessory or stimulator cells for 
      many lymphocyte responses. While knowledge of the response of other leucocytes 
      (e.g., lymphocytes, macrophages, and granulocytes) to freezing and thawing has 
      been established for some years, an understanding of the cryobiological 
      properties of DCs has not, hitherto, been determined specifically. Such 
      information is important both for establishing procedures for the long-term 
      storage of these cells for use in immunological procedures and for defining 
      freezing conditions that might selectively kill DCs in attempts to modulate the 
      immunogenicity of transplantable tissues during cryopreservation. Preparations of 
      rat and human spleen cells enriched for DCs were frozen to -60 degrees C at one 
      of six cooling rates (0.3, 1.5, 10, 20, 70, or 150 degrees C/min) using a 
      procedure that was established for pancreatic islets with 2 M dimethyl sulfoxide 
      (Me2SO) as the cryoprotectant. Following storage at -196 degrees C the survival 
      of thawed cells was assessed by evaluating both the numbers of cells recovered 
      after the complete process and the membrane integrity of the recovered cells 
      using a supravital fluorescent probe assay. Survival profiles for DCs showed a 
      dependence upon cooling rate similar to other lymphoid cells but DCs were more 
      sensitive to freezing injury than either lymphocytes or macrophages: Optimum 
      survival (75% recovery of numbers and 57% membrane integrity) of rat DCs was 
      achieved by slow cooling (0.3 degrees C/min). Optimal recovery of human DCs was 
      significantly higher (83% recovery of numbers and 72% membrane integrity) after 
      cooling at either 0.3 or 1.5 degrees C/min. The viable yield of DCs from both 
      species declined abruptly as cooling rate was increased, with less than 10% 
      survival after cooling at 20 degrees C/min and negligible survival after cooling 
      at 70 degrees C/min or greater. Analysis of variance of the survival data showed 
      that the response of DCs to freezing and thawing was significantly different (P 
      less than 0.005) from that of either lymphocytes or macrophages, thus providing 
      additional evidence that DCs are distinct from other leucocytes, especially 
      macrophages. This study defines conditions that either will provide effective 
      cryopreservation of DCs for immunological purposes or are most likely to bring 
      about their inactivation in cryobiological approaches to modulating tissue 
      immunogenicity.
FAU - Taylor, M J
AU  - Taylor MJ
AD  - MRC Medical Cryobiology Group, University Department of Surgery, Cambridge, 
      England.
FAU - London, N J
AU  - London NJ
FAU - Thirdborough, S M
AU  - Thirdborough SM
FAU - Lake, S P
AU  - Lake SP
FAU - James, R F
AU  - James RF
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - Netherlands
TA  - Cryobiology
JT  - Cryobiology
JID - 0006252
RN  - 0 (Cryoprotective Agents)
RN  - 0 (Fluorescent Dyes)
SB  - IM
MH  - Animals
MH  - Cell Membrane/ultrastructure
MH  - Cell Survival
MH  - *Cryopreservation/methods
MH  - Cryoprotective Agents
MH  - *Dendritic Cells/ultrastructure
MH  - Fluorescent Dyes
MH  - Humans
MH  - Leukocytes
MH  - Rats
MH  - Spleen/cytology
EDAT- 1990/06/01 00:00
MHDA- 1990/06/01 00:01
CRDT- 1990/06/01 00:00
PHST- 1990/06/01 00:00 [pubmed]
PHST- 1990/06/01 00:01 [medline]
PHST- 1990/06/01 00:00 [entrez]
AID - 0011-2240(90)90026-Z [pii]
AID - 10.1016/0011-2240(90)90026-z [doi]
PST - ppublish
SO  - Cryobiology. 1990 Jun;27(3):269-78. doi: 10.1016/0011-2240(90)90026-z.