91145 - HT098 - Colon - Frozen
Homo sapiens
Protocols: Enteroids were isolated from Matrigel droplets and manually sorted to remove dead or unhealthy entroids. Multiple healthy whole enteroids were pooled for each sample. Whole isolated crypts were embedded in 50uL Matrigel droplets in wells of a 24-well plate with growth media changed every 48 hours. LWRN Growth Media was produced as previously described (Miyoshi and Stappenbeck 2013). Conditioned media from L-WRN cells containing Wnt3a, Rspondin3, and Noggin, is mixed 1:1 with a 2x basal media comprised of: Advanced DMEM/F12 - 214mL, Glutamax (100x, 200mM) - 5mL, HEPES (100x, 1M) - 5mL, N2 Supplement (100x) - 5mL, B27 Supplement (50x) - 10mL, Pen/Strep (100x) - 5mL, N-acetylcystine (500 mM) - 1mL, Nicotinamide (1M) - 5mL. Frozen samples were derived from enteroids grown from cryporeserved human gastrointestinal biopsies. Fresh samples were derived of biopsy portions that were never frozen. RNA was isolated using either standard MagMAX or Qiagen RNeasy micro kits. Libraries were created with Takara SMARTer Stranded Total RNA Sample Prep Kits.
ENA first public
2018-06-30
ENA last update
2018-02-21
External Id
SAMEA104605964
INSDC center alias
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA
INSDC center name
Department of Internal Medicine, University of Michigan Medical School, Ann Arbor, MI 48109, USA
INSDC first public
2018-06-30T17:02:45Z
INSDC last update
2018-02-21T15:57:35Z
INSDC status
public
Submitter Id
E-MTAB-6516:91145 - HT098 - Colon - Frozen
age
70
broker name
ArrayExpress
common name
human
disease
normal
growth condition
organoid culture
individual
HT98
organism part
colon
sample name
E-MTAB-6516:91145 - HT098 - Colon - Frozen
sex
female
specimen with known storage state
frozen specimen