cell_suspension_17
Homo sapiens
Protocols: The human embryonic samples were collected from medical abortions. The samples were stored in hibernate E medium (Thermo Fisher A1370601) and transported to the research lab for dissection. The whole limb bud or upper part of the lower limbs were then dissected are stored in ice-cold PBS before single-cell isolation. Samples were chopped into a mash using a scalpel and incubated in 5ml of 2.5mg/ml liberase at 37 degrees for 30-40 min till fully digestion. Cells were spun down at 4C 750 for 5min and resuspended in 3ml of 1X RBC lysis buffer (eBioscience 00-4333, or diluted from eBioscience 00-4300) followed by 4-5 min incubation at RT. 10ml of 1X PBS was then added. The cells were then spun down at 4C at 750 for 5min, resuspended in 5ml of 2% FBS in 1X PBS, filtered through a 40um strainer, spun down again iat 4C 750g for 5min, resuspended in 50-200ul of 2% FBS in PBS, and spun down again. Cells were loaded according to the standard protocol of the Chromium single cell 5' V1 kit
disease
normal
individual
BRC2254
organism
Homo sapiens
organism
Homo sapiens
collection date
not collected
sex
male
scientific_name
Homo sapiens
common name
human
organism part
hindlimb
developmental stage
embryo stage
sampling site
isolate
not applicable
age
5.6
geographic location (country and/or sea)
not collected