Spermatogonia stem cell culture - (Jan/27/2008 )

Hello everyone,
I am going to do SSC culture. The ingredients of the medium are so complex. Is there easy way to make the medium? Thank for any helps.

Culture medium for the testis cells was StemPro-34
SFM (Invitrogen) supplemented with StemPro supplement (Invitrogen), 25
mg/ml insulin, 100 mg/ml transferrin, 60 mM putrescine, 30 nM sodium
selenite, 6 mg/ml D-(1)-glucose, 30 mg/ml pyruvic acid, 1 ml/ml DL-lactic
acid (Sigma), 5 mg/ml bovine albumin (ICN Biomedicals, Irvine, CA), 2
mM L-glutamine, 5 3 1025 M 2-mercaptoethanol, minimal essential medium
(MEM) vitamin solution (Invitrogen), MEM nonessential amino acid
solution (Invitrogen), 1024 M ascorbic acid, 10 mg/ml d-biotin, 30 ng/ml
b-estradiol, 60 ng/ml progesterone (Sigma), 20 ng/ml mouse epidermal
growth factor (Becton Dickinson, Bedford, MA), 10 ng/ml human basic
fibroblast growth factor (Becton Dickinson), 103 U/ml ESGRO (murine
leukemia inhibitory factor; Invitrogen), 10 ng/ml recombinant rat glial cell
line-derived neurotrophic factor (GDNF) (R&D Systems, Minneapolis,
MN) and 1% fetal calf serum (JRH Biosciences, Lenexa, KS).
BIOLOGY OF REPRODUCTION 69, 612–616 (2003)

quote]
I have the same research topic on spermatogonial stem cells. My study results show that those growth factors mentioned above are all very important to SSCs culture in vitro.
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Thank you Popsy,
I have not started to culture SSC. Do you use feeder layer? If yes, what kind do you use?

I use MEF as feeder cell in our culture systerm, what kind do you use? or try some no-feeder culture systerm?

I use MEF as feeder cell in our culture systerm, what kind do you use? or try some no-feeder culture systerm?