PMID- 23306781
OWN - NLM
STAT- PubMed-not-MEDLINE
DCOM- 20131025
LR  - 20220410
IS  - 0920-9069 (Print)
IS  - 1573-0778 (Electronic)
IS  - 0920-9069 (Linking)
VI  - 65
IP  - 5
DP  - 2013 Oct
TI  - Optimization of human umbilical cord mesenchymal stem cell isolation and culture 
      methods.
PG  - 819-27
LID - 10.1007/s10616-012-9528-0 [doi]
AB  - Human umbilical cord mesenchymal stem cells (hUCMSCs) are considered to be an 
      ideal replacement for bone marrow MSCs. However, up to date, there is no 
      convenient and efficient method for hUCMSC isolation and culture. The present 
      study was carried out to explore the modified enzyme digestion for hUCMSC in 
      vitro. Conventional enzyme digestion, modified enzyme digestion, and tissue 
      explant were used on hUCMSCs to compare their efficiencies of isolation and 
      culture, to observe primary cell growth and cell subculture. The results show 
      that the cells cultured using the tissue explant method had a longer culture 
      cycle (P < 0.01) and lower yield of primary cells per centimetre of umbilical 
      cord (P < 0.01) compared with the two enzyme digestion methods. Subculture 
      adherence and cell doubling took significantly less time with the tissue explant 
      method (P < 0.05) than with the conventional enzyme digestion method; however, 
      there was no significant difference between the tissue explant method and the 
      modified enzyme digestion method (P > 0.05). Comparing two enzyme digestion 
      methods, the modified method yielded more cells than did the conventional method 
      (P < 0.01), and primary cell adherence took significantly less time with the 
      modified method than with the conventional method (P < 0.05). Cell cycle analysis 
      of the third-generation hUCMSCs cultured by modified enzyme digestion method 
      indicated that most cells were quiescent. Immunofluorescence staining showed that 
      these cells expressed MSC markers CD44 and CD90. And Von Kossa and oil red O 
      staining detection showed that they could be differentiated into osteoblasts and 
      adipocytes with induction medium in vitro. This study suggests that hUCMSC 
      isolation and culture using 0.2 % collagenase II at 37  degrees C for digestion of 
      16-20 h is an effective and simple modified enzyme digestion method.
FAU - Han, Yan-Fu
AU  - Han YF
AD  - Department of Plastic Surgery, Affiliated Beijing Shijitan Hospital, Capital 
      Medical University, Beijing, 100038, People's Republic of China.
FAU - Tao, Ran
AU  - Tao R
FAU - Sun, Tian-Jun
AU  - Sun TJ
FAU - Chai, Jia-Ke
AU  - Chai JK
FAU - Xu, Guang
AU  - Xu G
FAU - Liu, Jing
AU  - Liu J
LA  - eng
PT  - Journal Article
DEP - 20130111
PL  - United States
TA  - Cytotechnology
JT  - Cytotechnology
JID - 8807027
PMC - PMC3967601
EDAT- 2013/01/12 06:00
MHDA- 2013/01/12 06:01
PMCR- 2014/10/01
CRDT- 2013/01/12 06:00
PHST- 2012/08/24 00:00 [received]
PHST- 2012/12/17 00:00 [accepted]
PHST- 2013/01/12 06:00 [entrez]
PHST- 2013/01/12 06:00 [pubmed]
PHST- 2013/01/12 06:01 [medline]
PHST- 2014/10/01 00:00 [pmc-release]
AID - 9528 [pii]
AID - 10.1007/s10616-012-9528-0 [doi]
PST - ppublish
SO  - Cytotechnology. 2013 Oct;65(5):819-27. doi: 10.1007/s10616-012-9528-0. Epub 2013 
      Jan 11.