PMID- 27323101
OWN - NLM
STAT- MEDLINE
DCOM- 20170210
LR  - 20170210
IS  - 1676-5680 (Electronic)
IS  - 1676-5680 (Linking)
VI  - 15
IP  - 2
DP  - 2016 May 25
TI  - Application of retrograde dissection method for isolation of bone marrow cells 
      from rat femurs and tibiae.
LID - 10.4238/gmr.15028178 [doi]
AB  - Currently, there is no practical and efficient method for the isolation of bone 
      marrow cells (BMCs) from rat femurs and tibiae. Here, we attempted to develop a 
      rapid, simple, effective, and non-contaminating method for the isolation of BMCs 
      from rat femurs and tibiae. Rat femurs and tibiae were dissected from the ankle 
      to the hip joint; subsequently, a three-step "locate-slide-twist" procedure was 
      performed using scissors and forceps to remove the femurs and tibiae completely, 
      from the surrounding musculature. The bones were flushed with phosphate-buffered 
      saline to harvest BMCs. The femurs and tibiae were dissected in 1.8 +/- 0.6 min, 
      and the BMC suspension preparation time was 13.1 +/- 2.3 min. The bone marrow 
      cavities did not incur any fractures or injuries during the isolation. Culture of 
      harvested BMCs for 72 h led to a significant increase in cell number from 4.4 +/- 
      0.3 x 106 to 6.9 +/- 0.7 x 10(6) (P < 0.01) with no significant decrease in 
      viability (98.1 +/- 0.6% vs 96.2 +/- 1.1%; P > 0.05). Microscopic examination of the 
      isolated BMCs after the 72-h incubation period revealed the no-microbial or 
      muscle cell contamination. Furthermore, flow cytometry revealed that cultured 
      BMCs (72-h culture) grew well. Here, we have reported a rapid, simple, effective, 
      and non-contaminating method for the isolation of BMCs from rat femurs and tibiae 
      by using retrograde dissection. This method can be used to harvest a large number 
      of viable BMCs without the risk of contamination from muscle and connective 
      tissues.
FAU - Li, C M
AU  - Li CM
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
FAU - Fu, B M
AU  - Fu BM
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
FAU - Zhang, L C
AU  - Zhang LC
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
FAU - Tang, B
AU  - Tang B
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
FAU - Zhu, L
AU  - Zhu L
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
FAU - Zhao, Y
AU  - Zhao Y
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
FAU - Zhang, J
AU  - Zhang J
AD  - Hepatobiliary and Pancreatic Surgery Ward, Second Affiliated Hospital of Kunming 
      Medical University, Kunming, China.
LA  - eng
PT  - Journal Article
DEP - 20160525
PL  - Brazil
TA  - Genet Mol Res
JT  - Genetics and molecular research : GMR
JID - 101169387
SB  - IM
MH  - Animals
MH  - Bone Marrow Cells/*cytology
MH  - Cell Culture Techniques/*methods
MH  - Cell Separation/*methods
MH  - *Cell Survival
MH  - Disease Models, Animal
MH  - Femur/*cytology
MH  - Rats
MH  - Tibia/cytology
EDAT- 2016/06/21 06:00
MHDA- 2017/02/12 06:00
CRDT- 2016/06/21 06:00
PHST- 2016/06/21 06:00 [entrez]
PHST- 2016/06/21 06:00 [pubmed]
PHST- 2017/02/12 06:00 [medline]
AID - gmr8178 [pii]
AID - 10.4238/gmr.15028178 [doi]
PST - epublish
SO  - Genet Mol Res. 2016 May 25;15(2). doi: 10.4238/gmr.15028178.