Frederick W. Kleinhans
Associate Professor, Emeritus

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Phone: 317-274-6900

Education

B.S., Physics, University of Michigan, Ann Arbor, MI 1965
Ph.D., Physics, Ohio State University, Columbus, OH, 1971

Research

My research is a collaborative effort with a number of biologists who are working on several cryobiological problems.  Cryobiology has seen wide use for efficient preservation of genetic mutants used in medical research (e.g. mice), assisted reproduction in humans and animals (e.g. the dairy industry), organ transplantation (e.g. pancreatic islet cells), and preservation of endangered species (e.g. the black-footed ferret). But no, not human heads or bodies.  We are trying to improve existing cryopreservation protocols and develop new ones for several cell types. Typically, successful cryopreservation requires the efficient removal of cell water and the introduction of cryoprotectants, e.g. glycerol, without osmotic damage or toxic injury. Thus it is useful to know the osmotic characteristics, water permeability (Lp), and solute permeability (Ps) of target cell membranes in order to model their response and optimize cryopreservation protocols. My work deals with understanding passive membrane transport, the design and analysis of membrane permeability experiments, and modeling cell responses.

Currently we are working on achieving or improving the cryopreservation of several species including zebrafish embryos and sperm, coral larvae, monkey sperm, oyster oocytes, and other marine species of aquaculture interest.   We are also investigating the fundamental mechanisms of cryo damage using mouse embryos and plant protoplasts.  Specifically, what event triggers the generally lethal formation of intracellular ice?  Cells typically do not freeze at their thermodynamic freezing point, but rather undergo supercooling which can reach ten’s of degrees C.  It is still not understood what drives intracellular nucleation of this super cooled water.

Current collaborators include Drs. Peter Mazur, University of TN, Knoxville; Mary Hagedorn, National Zoological Park, Washington, DC, and the Hawaii Institute of Marine Biology, Oahu, HI; Stuart Meyers, UC Davis, CA; Serean Adams, Cawthron Institute, New Zealand; and Keisuke Edashige and Magosaburo Kasai, Kochi University, Japan.  My work is funded via subcontracts with my collaborators who are currently funded by NIH, Friends of the (National) Zoo, and the Cawthron Institute.

Recent Publications (Reprints)

Kleinhans FW and Mazur P (2007)
Comparison of actual vs. synthesized ternary phase diagrams for solutes of cryobiological interest.
Cryobiology 54, 212-222..

Yamaji Y, Valdez DM Jr, Seki S, Yazawa K, Urakawa C, Jin B, Kasai M, Kleinhans FW and Edashige K (2006)
Cryoprotectant Permeability of Aquaporin-3 Expressed in Xenopus Oocytes.
Cryobiology 53 , 258-267.

Guenther JF, Sekia S, Kleinhans FW, Edashige K, Roberts DM, and Mazur P (2006)
Extra- and Intra-Cellular Ice Formation in Stage I and II Xenopus Laevis Oocytes.
Cryobiology 52 , 401-416.

Kleinhans FW, Guenther JF, Roberts DM and Mazur P(2006)
Analysis of Intracellular Ice Nucleation in Xenopus Oocytes by Differential Scanning Calorimetry.
Cryobiology 52 , 128-138.

Hagedorn M, Pan R, Cox EF, Hollingsworth L, Krupp D, Lewis TD, Leong JC, Mazur P, Rall WF, Macfarlane DR, Fahy G and Kleinhans FW (2006)
Coral Larvae Conservation: Physiology and Reproduction.
Cryobiology 52 , 33-47.

Mazur P, Seki S, Pinn IL, Kleinhans FW and Edashige K (2005)
Extra- and Intracellular Ice Formation in Mouse Oocytes.
Cryobiology 51, 29-53.

Mazur P, Pinn IL, Seki S, Kleinhans FW and Edashige K (2005)
Effects of Hold Time After Extracellular Ice Formation on Intracellular Freezing of Mouse Oocytes.
Cryobiology 51, 235-9.

Hagedorn M, Peterson A, Mazur P and Kleinhans FW (2004)
High Ice Nucleation Temperature of Zebrafish Embryos: Slow-Freezing is not an Option.
Cryobiology 49, 181-189.

Adams SL, Kleinhans FW, Mladenov PV and Hessian PA (2003)
Membrane Permeability Characteristics and Osmotic Tolerance Limits of Sea Urchin (Evechinus Chloroticus) Eggs.
Cryobiology 47, 1-13.

Edashige K, Yamaji Y, Kleinhans FW, and Kasai M (2003)
Artificial Expression of Aquaporin-3 Improves the Survival of Mouse Oocytes after Cryopreservation.
Biology of Reproduction 68, 87-94.

Edashige K, Yamaji Y, Kleinhans FW, and Magosaburo Kasai (2002)
Artificial Expression of Aquaporin-3 Improves the Survival of Mouse Oocytes after Cryopreservation. Biology of Reproduction2002, 10.1095/biolreprod.101.002394; Online.

Hagedorn M, Lance SL, Fonseca D, Kleinhans FW, Artimov D, Fleischer R, Hoque ATMS, Hamilton M and Pukazhenthi B (2002)
Altering Fish Embryos with Aquaporin-3: An Essential Step Toward Successful Cryopreservation.
Biology of Reproduction 67, 961-966.

 

 

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