Abstract：
:In cartilage tissue engineering, viable cell numbers should be correctly counted in the collagenase digest of the biopsied cartilage. However, this is a difficult task due to the presence of matrix debris, cell ghosts and their aggregates. To search for the correct cell counting method in this situation, we evaluated the utility of an automatic cell counting device, the NucleoCounter, and compared it with conventional staining using the LIVE/DEAD® kit. We first measured the cell numbers of a standard chondrocyte sample by the NucleoCounter, which showed a high accuracy (R(2) = 0.9999) and reproducibility (%CV: 2.00-8.66). We then calculated the cell numbers and viability in some collagenase digests of native cartilage using either the NucleoCounter or LIVE/DEAD® kit, revealing that the total cell numbers, viable ones and viability were highly correlated between them (R(2) = 0.9601, 0.9638 and 0.917, respectively). However, both the intrapersonal and interpersonal variabilities in the NucleoCounter was significantly decreased to about 1/20-1/5, compared to that of the LIVE/DEAD® kit. The NucleoCounter was regarded as a useful tool for simple, rapid, and highly reproducible cell counts, which may not only provide constant experimental data in a certain laboratory, but also contribute to the high reproducibility of the clinical results of cartilage tissue engineering among multiple institutions.
journal_name
Cytotechnologyjournal_title
Cytotechnologyauthors
Yonenaga K,Nishizawa S,Akizawa M,Asawa Y,Fujihara Y,Takato T,Hoshi Kdoi
10.1007/s10616-010-9304-ysubject
Has Abstractpub_date
2010-12-01 00:00:00pages
539-45issue
6eissn
0920-9069issn
1573-0778journal_volume
62pub_type
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