title: [Isolation and purification of human islet cells with semi-automated digestion].
abstract: OBJECTIVE
  To establish an semi-automated effective method for large-scale purification of islet cells from human pancreas.
  
  
  METHODS
  Human pancreas tissue was digested with collagenase P using a semi-automated pancreas-digestion system followed by purification in a HCA-Ficoll continuous gradient using Cobe2991 cell separator. After isolation, the islet cell yield and purity was evaluated with light microscope with DTZ staining, and the islet function assessed by insulin release assay in vitro.
  
  
  RESULTS
  The number of the islets collected from each pancreas averaged 38 6201-/+78 219 islet equivalents (IEQ) before purification, and 231 420-/+28 054 IEQ after the purification with discontinuous gradient centrifugation. From each gram of the pancreatic tissue, 3148-/+317 IEQ were obtained with an average purity of (62.81-/+2.68) %. The purified islets responded well to high-concentration (16.7 mmol/L) glucose stimulation with a 2.53-fold increase of insulin secretion over the basal level (3.3 mmol/l, P<0.001).
  
  
  CONCLUSIONS
  The established semi-automated method can be applicable for large-scale purification of fully functional islet cells from human pancreas.
authors.affiliation: Department of Endocrinology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. jimzhua@163.com
authors.forename: Hua
authors.initials: H
authors.lastname: Zhang
authors.forename: De-hong
authors.initials: DH
authors.lastname: Cai
authors.forename: Jun-ling
authors.initials: JL
authors.lastname: Han
authors.forename: Guo-hong
authors.initials: GH
authors.lastname: Wei
authors.forename: Jia
authors.initials: J
authors.lastname: Sun
authors.forename: Wei-ping
authors.initials: WP
authors.lastname: Sun
authors.forename: Jian
authors.initials: J
authors.lastname: Zhang
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Long-yi
authors.initials: LY
authors.lastname: Zeng
authors.forename: Jian-ping
authors.initials: JP
authors.lastname: Weng
mesh_terms: D002452:Cell Count; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods*; D002470:Cell Survival; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000378:metabolism; D000078790:Insulin Secretion; D007515:Islets of Langerhans / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015203:Reproducibility of Results
chemical_list: D007328:Insulin; D005947:Glucose