What are the best culturing media, supplements, and protocols for expansion of HSCs?
Dear Farshid,
I just briefly wanted to email you concerning your HSC culture question. In the attached review, you find a rather comprehensive overview over a) different supplements for HSC culture b) culture periods and c) subsequent assays that have been used to assess successful expansion, which you can hopefully tailor to your research question.
http://www.cell.com/trends/molecular-medicine/fulltext/S1471-4914(17)30110-7
A company that supposedly has superb media for HSC culture is Stemcell Diagnostics from Canada. My fav choice (that always worked like a charm) is StemMACS from Miltenyi Biotec.
https://www.stemcell.com/media/files/manual/MA28404-Human_Colony_Forming_Unit_Assays_Using_MethoCult.pdf
http://www.miltenyibiotec.com/en/products-and-services/macs-cell-culture-and-stimulation/media/stem-cell-media/stemmacs-hsc-cfu-media-human.aspx
Unfortunately, they both don't come cheap!
All the best & good luck with your experiment,
Michael
Stem cells technology offer a kit for HSCs Isolation with magnetic system, they also have supplements for growth. I use them and they are working perfectly fine.
Preferred Cell Systems offers the largest selection of assays, including ColonyGro, CAMEO-4, CAMEO-96, HALO, HemoFLUOR and HemoLIGHT for up to 8 different stem cell and numerous progenitor and precursor cell populations from 9 different species. Please visit the website at www.preferred-cell-systems.com.
Hello Farshid,
You could certainly use a methylcellulose CFU assay such as that suggested by others or the ColonyGro, CAMEO-4 or CAMEO-96, but if you have not had experience in counting colonies, you are going to get bogged down. The MethoCult products do not allow you to detect primitive stem cells because they do not contain TPO, which allows the megakaryopoietic lineage to be detected. The CFC-GEMM 1 ColonyGro, CAMEO-4 or CAMEO-96 assays include TPO. Having said that, MethoCult and other products are extremely insensitive when it comes to rare, multipotent stem cells. That is why we developed the instrument-based assays HALO, HemoFLUOR and HemoLIGHT. HALO is about 5,000 time more sensitive than any CFU assay.
Preferred Cell Systems offers both hematopoietic stem cell culture media and complete assay kits. www.preferred-cell-systems.com.
Hi Farshid Amiri
As an alternative to the colony assays with MethoCult as mentioned above, which allow you to quantify the number of hematopoietic progenitor cells in a sample - we also offer liquid culture conditions for optimal expansion of CD34+ cells isolated from a variety of tissues. Using our optimized conditions you can achieve 80 - 100X expansion of CD34+ cells isolated from cord blood using StemSpan SFEM II medium, CD34+ expansion cytokine cocktail and small molecule UM171.
Our optimized protocol is here:
https://www.stemcell.com/technical-resources/educational-materials/tech-tips/serum-free-expansion-of-cd34-cells-with-sfem-ii.html?utm_source=researchgate
Please let me know if you have any questions,
Amanda
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