See discussions, stats, and author profiles for this publication at: https://www.researchgate.net/publication/259687687 Setting Up an Andrology Laboratory Chapter · January 2014 CITATIONS READS 0 735 1 author: Ashok Agarwal Cleveland Clinic 1,844 PUBLICATIONS 31,897 CITATIONS SEE PROFILE Some of the authors of this publication are also working on these related projects: Antioxidant therapy in idiopathic oligoasthenoteratozoospermia View project Sperm DNA Fragmentation View project All content following this page was uploaded by Ashok Agarwal on 14 January 2014. The user has requested enhancement of the downloaded file. 16 Setting Up an Andrology Laboratory Kelly S Athayde, Ashok Agarwal ABSTRACT diseases or malfunctions related to male fertility. Utilizing both simple and complex techniques, its evaluation Male factor infertility is a sole or contributing factor in approximately 40% of the infertile couples and therefore provides useful information for the infertility specialist investigating the cause of the male infertility constitutes a in the management of the subfertile man. Besides major part of the diagnosis and treatment of infertile couples. diagnostic activities, it should provide therapeutic The andrology laboratory supports and conducts clinical procedures for patients requiring assistance in achieving assessment of the semen and helps in the diagnosis of pregnancy such as Assisted Reproduction techniques: various diseases and malfunctions related to male infertility. Development of advanced assisted reproductive Intrauterine Insemination (IUI) and In Vitro Fertilization techniques in the last decade has increased the scope of (IVF). The Andrology laboratory also works in parallel today’s andrology laboratories which are also involved in with endocrinology, genetic, and in vitro fertilization cryopreservation of gametes and reproductive tissues laboratories to diagnose and assist in the treatment of preserving the reproductive capacities of couples for various patients facing difficulties conceiving or seeking reasons. This chapter serves as a practical guide for setting-up an preservation of their reproductive capacity by andrology laboratory for diagnostic and therapeutic cryopreserving their gametes or reproductive tissue. purposes. This chapter will provide readers, in a very practical way, with information on the basic requirements for INTRODUCTION setting up and operating an Andrology laboratory for diagnostic and therapeutic activities. “Andrology” is the field of medicine that deals with matters affecting the male reproductive system. The LABORATORY ASPECTS AND PROCEDURES earliest use of this term appeared in 1891 in the Journal of American Medical Association when it reported on Numerous procedures can be performed in the the formation of the American Andrological Association.1 Andrology Laboratory. Due to different applications Because male infertility is the primary or contributing (clinical and research) and clinical correlations in the factor in more than 40% of infertile couples 2, this field current scientific literature, some of these procedures may has been the object of constant research and has therefore not be included in this chapter. seen advances in diagnostic tests and tools used for the Semen analysis is the most common test to be investigation of male infertility. conducted in the Andrology Laboratory. It reveals how The Andrology Laboratory encompasses clinical many spermatozoa are in the ejaculate, the vigor with laboratories under both Pathology and Laboratory which they move (quantitative motility), and how normal Medicine. It is entrusted with supporting clinical these cells look morphologically. Complementary assessments of semen that may help diagnose potential assessments should include viability, the presence of 158 ANDROLOGY LABORATORY MANUAL round cells and their identification (i.e. leukocytes or Azoospermia Screen Procedure immature sperm cells), and detection of antibodies. Antibody Assessment Additional tests can be performed based on the results Test for anti-sperm antibodies on the sperm surface obtained from this initial analysis. Such tests may Test for free antibodies, directed against the sperm surface complement the initial analysis and provide more (seminal plasma, serum or cervical mucus) information about sperm cell functionality and structure. Biochemical markers such as Reactive Oxygen Species Sperm DNA Damage Test (ROS) and tests that assess sperm DNA damage and the Sperm Chromatin Structure Assay (SCSA) inducibility of the Acrosome Reaction (AR) can provide Terminal deoxynucleotidyl transferase dUTP nick end a more specific functional assessment of these gametes. labeling (TUNEL) Bioassays that assess gamete interaction, such as the COMET, also called single cell gel electrophoresis (SCGE). sperm-hamster egg penetration assay (SPA) and the human sperm-zona pellucida binding test (hemizona Therapeutic assay, HZA), evaluate the occurrence of spontaneous Sperm Preparation for Intrauterine Insemination (IUI) sperm AR and the capacity for penetration, respectively. or In vitro Fertilization (IVF) They have strong diagnostic power but, like the AR test, Preparation of cryopreserved semen for IUI or IVF still require better standardization before they can be 3 introduced as clinical tools. Moreover, the bioassays Techniques require hamster eggs and non-fertilizable human oocytes, which can impose extra practical limitations for their Gradient method introduction into routine laboratory practice. Swim-up method The most commonly performed tests are listed below. Simple wash (concentration technique) They can be classified in two groups: diagnostic and Cryopreservation therapeutic. Semen Diagnostic Procedures Epididymal aspirate Testicular Tissue Complete Semen Analysis Macroscopic parameters (color, volume, pH, viscosity, LABORATORY DESIGN liquefaction, and agglutination) Before working on any laboratory design, local Microscopic parameters (sperm concentration, motility, regulations must be carefully studied. These regulations morphology, presence of round cells, and different types may differ between countries or even between states, and of debris) compliance is not optional. Ideally, the Andrology Laboratory should contain Detection of Leukocytes in Semen distinct areas for diagnostic and therapeutic procedures Myeloperoxidase test (Endtz) (Figure 16.1). Therapeutic procedures should be performed in a sterile environment, which can be Sperm Vitality and Membrane Viability achieved by using a laminar flow hood. The physical area should be determined according to a pre-estimation of Eosin-Nigrosin test the number of tests to be offered by the service and the Hypo-osmotic Swelling test (HOS) volume of procedures expected to be performed. This will Biochemical determine the number of technicians that must be employed and the different types of equipment that must Fructose qualitative and quantitative be made available such as microscopes, cell counters, incubators, etc. New laboratories may not have an Oxidative Stress established range of workload due to regional and market Reactive Oxygen Species (ROS) variations and thus should always keep in mind plans Total Antioxidant Capacity (TAC) for future expansion. SETTING UP AN ANDROLOGY LABORATORY 159 Figure 16.1: Andrology Laboratroy and Reproductive Tissue Bank – Cleveland Clinic, Cleveland, Ohio Even a small physical area can have its space optimized according to its disposition and organization. If the Figure 16.2: Example of liquid nitrogen containers for storage laboratory is going to offer cryopreservation, there should be space for the liquid nitrogen (lN2) containers that will store tissue samples (Figure 16.2). Due to the chemical properties of lN2, a laboratory should follow the proper safety guidelines for area ventilation and oxygen monitoring. Containers should be kept locked or in areas of restricted access. If containers are not checked daily, the laboratory should have alarms that sound when levels of lN2 fall too low. Semen Collection Room The semen collection room (SCR) is a very important aspect of an Andrology Laboratory. Therefore, it must be planned with attention to design, especially in regards to location. Usually the SCR is located in proximity to the laboratory. Places that provide privacy (avoid busy corridors, lobbies or reception areas) are appropriate. The room can be equipped with a love seat or a small sofa and be supplied as needed with clean towels and sheets Figure 16.3: Example of semen collection room (Figure 16.3). There should be a sink or a washing station 4 for use before and after collection. Not all collection rooms of media. The survey responders also rated videos as provide toilets and, if this is the case, a bathroom should the most helpful media followed by magazines, with the be available nearby. The furnishings must be kept simple preferred theme being heterosexual sex. In the same with preference for easy-to-clean materials or surfaces. study, the responders preferred instructions on collection Instructions for collection should be posted in the SCR technique to be given face-to-face rather than in writing. even when they are explained to the patients by the nurse Office and Break Room or laboratory technician. An interesting survey on patient satisfaction reported that the main areas of concern Clerical staff will also require office space. Patients’ charts regarding the SCR were: sound privacy of the room, and files need to be kept within easy access of the furnishing,