Method and Device for Removal of Cryoprotectant from Cryopreserved Biological Cells and Tissues

Novel methods and devices for removing cryoprotectant from cryoprotectant-containing liquids, and from cells residing therein, are disclosed. In one aspect, the method comprises passing the cryoprotectant-containing liquid through at least one semipermeable hollow fiber membrane contained in a hollow module in a first direction, while passing a liquid which is substantially free of cryoprotectant through the hollow module in a second direction to remove cryoprotectant across a diffusion gradient. In another aspect, a device is described for removing cryoprotectant from a liquid, comprising a hollow module with at least one semipermeable hollow fiber membrane therein for accomplishing such counter-current diffusion removal of cryprotectant. A software program is also provided for predicting optimal flow rates through the device of the invention, thereby allowing optimal cryoprotectant removal regardless of the cryoprotectant used or the material from which the semipermeable hollow fiber membrane is fabricated

Methods and Devices for Cryopreservation of Biological Cells and Tissues

Novel methods, compositions, and devices for achieving optimal cooling of living cells during cryopreservation are disclosed. In one aspect, the method comprises gradually cooling the cell to a first predetermined temperature, followed by rapidly cooling the cell to a second predetermined temperature. In another aspect, a device is described for achieving a desired cooling rate for a cell, comprising a first container for holding a cell, a second container for holding the first container, and optionally a frame for holding the first container in a spaced apart relationship with the second container. The method of the invention comprises placing cells into the first container, placing the first container in the second container and sealing the second container, and placing the second container in a suitable cooling device. In yet another aspect, novel cryoprotectant compositions are provided comprising conventional cryoprotectant plus one or more high molecular weight cryoprotectants

Assessment of Cryoprotectant Concentration by Electrical Conductivity Measurement and Its Applications in Cryopreservation

This chapter presents an important application of the electrical conductivity measurement in cryopreservation. Long-term cryopreservation of cells and tissues is essential in both clinical treatments and fundamental researches. In order to reduce the cryo-injury to the cells during cryopreservation, cryoprotective agents (CPAs) should be added before freezing, but also removed after thawing duo to the cytotoxicity. In these steps, severe osmotic stresses may result in injuries to the cells too. Therefore, monitoring the addition and removal of CPAs to the cell samples is critical in order to prevent the osmotic injury. In this chapter, the electrical conductivity measurement was applied to assess the CPA concentration in cryopreservation. Firstly, the standard correlations between the CPA concentration and the electrical conductivity of the solutions (including CPA-NaCl-water ternary solutions and CPA-albumin-NaCl-water quaternary solutions) were experimentally obtained for a few mostly used CPAs. Then a novel ?dilution-filtration? system with hollow fiber dialyzer was designed and applied to remove the CPA from the solutions effectively. Measurement of electrical conductivity was validated as a safer and easier way to on-line and real-time monitoring of CPA concentration in cell suspensions. This work demonstrated a very important application of electrical conductivity in the biomedical engineering field

Progress in Cryopreservation of Stem Cells and Immune Cells for Cytotherapy

Cellular therapy with stem and immune cells has demonstrated significant success both in clinical treatments and the industrial market. Cryopreservation is a necessary and essential component of cellular therapy. In this chapter, first of all, some basic theories of cryoinjury and techniques in cryopreservation are reviewed. Then it focuses on the progress of cryopreservation of stem cells and immune cells, including new protocols and techniques, alternative cryoprotective agents (CPA), side effects after transplantation, and advances in reducing adverse reactions. Strategies to minimize adverse effects include medication before and after transplantation, optimizing the infusion procedure, reducing the CPA concentration or using alternative CPAs for cryopreservation, and removing CPA prior to infusion. Traditional and newly developed approaches including methods and devices for CPA removal are discussed. Future work is recommended including further optimization of cryopreservation protocols especially for lymphocytes; standardization of the optimized protocols with temperature monitoring and quality control; exploration of DMSO-free, serum-free, and even xeno-free media for cryopreservation; development of simple, reliable, and cost-effective devices for cryopreservation; and more fundamental cryobiological studies to avoid cellular injury.Keywords: cryopreservation, stem cell, immune cell, cytotherap

Research progress on GlnR-mediated regulation in Actinomycetes

This review constitutes a summary of current knowledge on GlnR, a global regulator, that assumes a critical function in the regulation of nitrogen metabolism of Actinomycetes. In cross-regulation with other regulators, GlnR was also shown to play a role in the regulation of carbon and phosphate metabolisms as well as of secondary metabolism. A description of the structure of the GlnR protein and of its binding sites in various genes promoters regions is also provided. This review thus provides a global understanding of the critical function played by GlnR in the regulation of primary and secondary metabolism in Actinomycetes

Cryobiology and Cryopreservation of Sperm

Low temperature has been utilized to keep living cells and tissues dormant but potentially alive for cryopreservation and biobanking with great impacts on scientific and biomedical applications. However, there is a critical contradiction between the purpose of the cryopreservation and experimental findings: the cryopreserved cells and tissues can be fatally damaged by the cryopreservation process itself. Contrary to popular belief, the challenge to the life of living cells and tissues during the cryopreservation is not their ability to endure storage at cryogenic temperatures (below −190°C); rather it is the lethality associated with mass and energy transport within an intermediate zone of low temperature (−15 to −130°C) that a cell must traverse twice, once during cooling and once during warming. This chapter will focus on (1) the mechanisms of cryoinjury and cryopretection of human sperm in cryopreservation, and (2) cryopreservation techniques and methods developed based on the understanding of the above mechanisms

Development of a Novel Electromagnetic Rewarming Technology for the Cryopreservation of Stem Cells with Large Volume

Applications of stem cells have been playing significant roles in scientific and clinical settings in the last few decades. The foundation of these approaches is successful cryopreservation of stem cells for future use. However, so far we can only cryopreserve stem cell suspension of small volumes in the order of 1 mL mostly due to the lack of an effective rewarming technique. Rapid and uniform rewarming has been approved to be beneficial, and sometimes, indispensable for the survival of cryopreserved stem cells, inhibiting ice recrystallization or devitrification. Unfortunately, the conventional water bath thawing method failed in providing the rapid and uniform rewarming. The conversion of electromagnetic (EM) energy into heat provides a possible solution to this problem. This chapter will focus on (1) analysis of the combined EM and heat transfer phenomenon in the rewarming of a biospecimen, (2) numerical investigation of the rewarming system, (3) practical setup of an EM resonance system, and (4) test of heating performance with large volume of cells