Genekam Stem cell Isolator (sterile) 250ml

Cat. No.	Name of ready to use PCR Kit	Storage	DOC	Amount	Price*
STEM14	Genekam Stem cell Isolator (sterile) 250ml			250ml	150,-€

Isolation Solution: Genekam Stem Cell Isolater*

Genekam Stem cells isolater from Umbilical blood samples: Genekam Biotechnology AG has developed a simple, economical method to isolate stem cells from umblical cord. You just add the solution into umblical blood and centrifuge it. You can isolate the stem cells from small volumes of fluid also e.g. 100 µl (provided they contain the stem cells) as well as larger size samples. Now you can isolate the stem cells with different methods as well as you can do further analysis. This method is far better than density gradient method to isolate the stem cells.
Genekam Stem cell (SC) isolator from umbilical cord blood is ready to use sterile solution for isolating stem cells containing populations (SCP) quickly from blood, buffy coats, bone marrow aspirates etc of different mammals like human, rat, primates, cattle, sheep, goat, pig, horse, camel, dog, cat etc.

This kit contains followings:

SC isolator
Washing buffer
Here are a few examples of application of SC isolater:

A. Isolation of SCP from umbilical cord blood:

Add 900 µl of Genekam SC isolater to each tube containing up to 100 µl of umbilical cord blood in one sterile tube as this is ration (1:10) of solution to the sample e.g. if you want to isolate cells from 4 ml blood of and add 36 ml of SC isolator. You have to decide howmuch blood sample do you want to use to isolate SCP as you will get a lot of SCP against the conventional method of density gradient centrifugation.
Warning: Kindly work with smallest volume of sample to gain some experience with this kit instead of destroying whole sample for first time.
Gently vortex each tube immediately after adding the solution. Keep at room temperature, protected from light, for 7-10 minutes.
Centrifuge 1600 rpm for 5 minutes. Aspirate or discard supernatant without disturbing pellet.
Resuspend the pellet in washing buffer and wash 2X while centrifuging for 1600 rmp with brake for 5 minutes each.
Resuspend the pellet and proceed with further procedures e.g. counting the cell number and go for your isolation of stem cells e.g. cell culturing or magnetic beads.
Stem cells can be isolated through pouring the isolated SCP (1x 106/cm2 suspended in DMEM with 10-15% FCS or other media; it is recommended to use FCS precoated plates as they may increase the chances to get the stem cells) into cell culture plates. Keep them at 37°C and 5% for 24-72 hours. After that discard the supernant as the stem cells will be adherent on plastic as adherent fibroblastiod cells, they can be grown and subcultured. Subconfluence of these cells can be isolated with trypsin. Usually cells at the second passage and thereafter can be replated 3.5 to 5 x 103/cm2.

B. Isolation of SCP from human bone marrow aspirate:

Add 400 µl of Genekam SC isolater to each tube containing up to 200 µl of human bone marrow in one sterile tube as this is ration of solution to the sample (1:3 ration) e.g. if you want to isolate SCP from 4 ml of bone marrow aspirate and add 8 ml of SCP isolator. You have to decide about the volume of bone marrow aspirate you want to use to isolate the cells because you will get a lot of cells against the conventional method of density gradient centrifugation.
Warning: If you donot have any experience with this kit, kindly use smallest volume of sample to have experience with this kit before you destroyed the whole sample. You may have to adjust ration e.g. you have still SCP containing some erythrocytes at the end, you have to use more SC isolator (1 volume sample and 4 volume SC isolator e.g. 200 µl sample and 600 µl of SC isolator) or wash one time more. Usually the above said protocoal in point 1 is good enough to isoate.<
Gently vortex each tube immediately after adding solution. Keep at room temperature, protected from light, for 7-10 minutes.
Centrifuge 1600 rpm for 5 minutes. Aspirate or discard supernatant without disturbing pellet.
Resuspend the pellet in washing buffer and wash 2X while centrifuging for 1600 rmp with brake for 5 minutes each.
Resuspend the pellet and proceed with further procedures e.g. counting the cell number and isolating the samples with different methods e.g. cell culturing or magnetic bead separation.
Stem cells can be isolated through pouring the isolated SCP (1x 106/cm2 suspended in DMEM with 10-15% FCS or other media; it is recommended to use FCS precoated plates as they may increase the chances to get the stem cells) into cell culture plates. Keep them at 37°C and 5% for 24-72 hours. After that discard the supernant as the stem cells will be adherent on plastic as adherent fibroblastiod cells, they can be grown and subcultured. Subconfluence of these cells can be isolated with trypsin. Usually cells at the second passage and thereafter can be replated 3.5 to 5 x 103/cm2.

* only for research use.