SpermActive

GM501 SpermActiveSperm Air edited no CE mark

  • Ready to use
  • HEPES (15mM) and bicarbonate buffered
  • Suitable for washing, swim-up and density gradient centrifugation
  • For handling of testicular tissue
  • Contains Gentamicin (10 mg/liter) and Phenol Red
  • Includes highest available purity (Ph. Eu./USP) Human Serum Albumin (5.00 g/liter)
  • Can be used safely for up to 7 days after opening when stored at 2 - 8°C and sterile conditions are maintained
  • A Certificate of Analysis for each batch is available on hamiltonthorne.com.
  • Available in 20ml and 50ml sizes

GM501 SpermActive Intended Use

GM501 SpermActive is intended for the handling and preparation of sperm for use in assisted reproduction procedures. GM501 SpermActive is not intended for use in intrauterine insemination (IUI) procedures.

GM501 SpermActive is a ready-to-use medium designed for sperm washing, swim-up techniques and density gradient centrifugation, as well as for testicular tissue.

GM501 SpermActive Instructions for Use

  • Prior to use, GM501 SpermActive must be equilibrated overnight in a humidified CO2-incubator (at 5 - 7% CO2 , 37°C).
  • Any laboratory procedures described herein are recommendations only. Each laboratory must establish and validate its own procedures for preparation and use.

GM501 SpermActive Composition

  • NaCl, KCl, KH2PO4, MgSO, CaCl2
  • Bicarbonate, HEPES, EDTA
  • Glucose, Lactate, Pyruvate
  • Non-essential and essential Amino Acids, Alanyl-Glutamine
  • Human Serum Albumin, Gentamicin, Phenol Red

GM501 SpermActive Specifications

pH (at 37°C, 6%CO2) 7.20 - 7.50
Osmolality (mOsm/kg) 270 - 290
Sterility (sterile/SAL 10-3)  No Growth
Endotoxins (LAL, EU/ml) < 0.25
Human Sperm Survival Assy (HSSA) ≥ 80% of Control Motility at 24h

GM501 SpermActive Order Codes

Part # Size Storage Shelf Life*
4 GM501SA-020  1 x 20ml 2 - 8°C 6 months
4 GM501SA-050  1 x 50ml 2 - 8°C 6 months
    * From the date of manufacture
 
MKT-2011 Rev. A, 06-Nov.-2020 
 
 
Attachments:
FileDescriptionFile size
Download this file (MKT-2011-GM501-SpermActive-RevA-Nov-06-2020-Final.pdf)MKT-2011 GM501 SpermActive Rev.AGM501 SpermActive Brochure1806 kB

Direct Swim-up Protocol

Media

The below protocol reflects the use of GM501 SpermAir or GM501 SpermActive for Direct Swim-up.

  1. Transfer 2 ml of  warmed GM501 SpermActive or GM501 SpermAir into a new conical tube (Fig. 1).
  2. Underlay with 1 ml of liquified semen (Fig. 2).
  3. Place cap on tube but do not tighten. Place in CO2 incubator for 1 hour (GM501 SpermActive) or warm to 37°C for 1 hour (Fig. 3).

Washing

  1. Without dispersing the lower layer, aspirate the upper media layer containing the motile sperm and transfer to a conical centrifuge tube (Fig. 4).
  2. Centrifuge tube at 300-400x g for 6 minutes (Fig. 5). (See below on how to calculate g-force)
  3. Without dispersing the pellet, aspirate and discard the supernatant (Fig. 6).
  4. Resuspend the pellet in 1ml of warmed GM501 SpermAir medium (Fig. 7).
  5. Centrifuge at 300-400x g for 6 minutes (Fig.8).
  6. Without dispersing the pellet, aspirate and discard the supernatant (Fig. 9).
  7. Repeat Steps 4 through 6 (Fig. 7 - 9).
  8. For IVF: Resuspend the pellet in 0.1-1.0 ml of equilibrated culture medium. For ICSI or Insemination: Resuspend the pellet in warmed GM501 SpermAir or equilibrated GM501 SpermActive.

 

direct swimup

Indirect Swim-up Protocol

Media

The below protocol the use of GM501 SpermAir or GM501 SpermActive for Indirect Swim-up.

Washing 1

  1. Transfer 5 ml of sperm washing media into a conical centrifuge tube.
  2. Add 1.0 to 3.0 ml liquified semen and mix well (FIg. 1).
  3. Centrifuge at 300-400x g for 10 minutes (Fig.2). (See below on how to calculate g-force)
  4. Aspirate the supernatant and discard (Fig. 3).

Swim-up

  1. Overlay the pellet with 2ml of equilibrated GM501 SpermActive or warmed GM501 SpermAir medium (Fig. 4).
  2. Place cap on tube but do not tighten and place in CO2 incubator (GM501 SpermActive) or warm to 37°C (GM501 SpermAir) for 1 hour (Fig. 5).
  3. Aspirate the supernatant containing the motile sperm and transfer to a new tube (Fig. 6).

Washing 2

  1. Add 1 ml of equilibrated GM501 SpermActive or warmed GM501 SpermAir medium to the aspirated supernatant (Fig.7).
  2. Centrifuge tube at 300-400x g for 6 minutes (Fig. 8).
  3. Without dispersing the pellet, aspirate and discard the supernatant (Fig. 9).
  4. Repeat Steps 1 through 3 (Fig. 7-9).
  5. For IVF: Resuspend the pellet in 0.1-1.0 ml of equilibrated culture medium. For ICSI or Insemination: Resuspend the pellet in warmed GM501 SpermAir or equilibrated GM501 SpermActive. 

 

direct swimup

G-force or RPM of Centrifuge

Calculation of G-force or RPM of Centrifuge

The g-force (g) of your centrifuge can be calculated using this formula:

     g = 0.0000284 x r x rpm2
     where:
            r = radius of centrifuge rotor (inches)
            rpm = revolutions per minute

     Example:

            r = 5 inches
            rpm = 1600
            g = 0.0000284 x 12 x 11002 = 364 g

If the g-force (g) is known, the rpm can be calculated using this formula:

      rpm = Square root {g/ (0.0000284 x r)} = rpm

      Example:

             r = 5 inches
            g = 375
            rpm = Square root {400/(0.0000284 x 12)] = 1,625 rpm

Please refer to your centrifuge’s instructions for use for more information.