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Primary culture of chondrocytes after collagenase IA or II treatment of articular cartilage from elderly patients undergoing arthroplasty Cover

Primary culture of chondrocytes after collagenase IA or II treatment of articular cartilage from elderly patients undergoing arthroplasty

Open Access
|Apr 2021

Figures & Tables

Figure 1

Comparison of cell yield between 3 chondrocyte isolation methods. Chondrocytes were isolated from elderly human articular cartilage using 3 methods. We divided cartilage slices in 3 groups: Group I, digested sequentially with pronase E and collagenase IA; Group II, digested with collagenase II only; or Group III, digested sequentially with pronase E and collagenase II. After digestion, total cell number was counted and mean cell yield (No. of chondrocytes per mg wet weight of cartilage) is plotted. We compared the difference in cell yield between Groups I and II with a 2-tailed t test assuming unequal variances. Error bars represent SD. SD, standard deviation.
Comparison of cell yield between 3 chondrocyte isolation methods. Chondrocytes were isolated from elderly human articular cartilage using 3 methods. We divided cartilage slices in 3 groups: Group I, digested sequentially with pronase E and collagenase IA; Group II, digested with collagenase II only; or Group III, digested sequentially with pronase E and collagenase II. After digestion, total cell number was counted and mean cell yield (No. of chondrocytes per mg wet weight of cartilage) is plotted. We compared the difference in cell yield between Groups I and II with a 2-tailed t test assuming unequal variances. Error bars represent SD. SD, standard deviation.

Figure 2

Viability of chondrocytes isolated from 11 cartilage specimens using pronase E and collagenase IA. Cartilage specimens obtained from 11 patients were first incubated with 0.4% pronase E and then with 0.02% collagenase IA. The viability of chondrocytes isolated from each patient is plotted. The long dashed line indicates the mean level of chondrocyte viability (84%) computed from 11 specimens. The short dashed lines indicate one standard deviation from the mean (8%).
Viability of chondrocytes isolated from 11 cartilage specimens using pronase E and collagenase IA. Cartilage specimens obtained from 11 patients were first incubated with 0.4% pronase E and then with 0.02% collagenase IA. The viability of chondrocytes isolated from each patient is plotted. The long dashed line indicates the mean level of chondrocyte viability (84%) computed from 11 specimens. The short dashed lines indicate one standard deviation from the mean (8%).

Figure 3

Representative light microscopic images of monolayer human chondrocytes. (A) Passage 0 human chondrocytes isolated with pronase E and collagenase IA seeded at 2.5 × 105 cells per cm2 and visualized at day 2 post-seeding (4× objective lens, scale bar 200 μm) and (B) detail of the insert in the black square as imaged with a 10× objective (scale bar 50μm). (C) Passage 0 chondrocytes at day 10 post-seeding imaged with a 4× objective (scale bar 200 μm). (D) Passage 1 chondrocytes at day 1 post-seeding imaged with a 10× objective (scale bar 200 μm).
Representative light microscopic images of monolayer human chondrocytes. (A) Passage 0 human chondrocytes isolated with pronase E and collagenase IA seeded at 2.5 × 105 cells per cm2 and visualized at day 2 post-seeding (4× objective lens, scale bar 200 μm) and (B) detail of the insert in the black square as imaged with a 10× objective (scale bar 50μm). (C) Passage 0 chondrocytes at day 10 post-seeding imaged with a 4× objective (scale bar 200 μm). (D) Passage 1 chondrocytes at day 1 post-seeding imaged with a 10× objective (scale bar 200 μm).

Donor patient characteristics and wet weights of cartilage harvested

Patient characteristicNo. of patients% of total patientsMean yield of wet cartilage ± SD (mg)
Age (y)50–59163430
60–696333760 ± 1489
70–798444211 ± 1716
80–893172740 ± 1221
SexMale6333708 ± 1035
Female12673804 ± 1781
DiagnosisFemoral neck fracture11613245 ± 1137
Femoral head necrosis4223253 ± 765
Hip osteoarthritis166450
Knee osteoarthritis2116370

Donor patient characteristics and chondrocyte yield for each treatment group

TreatmentCharacteristicNo. of patientsChondrocyte yield (mean cells ±SD)
Group I (n = 11): pronase E + collagenase IAAge (y)60–6942940 ± 629
70–7962390 ± 1050
80–8912130
SexMale32514 ± 601
Female82586 ± 992
DiagnosisFemoral neck fracture62587 ± 1056
Femoral head necrosis23002
Hip osteoarthritis13030
Knee osteoarthritis21837

Group II (n = 5): collagenase IIAge (y)50–5912729
60–691747
70–791669
80–892789
SexMale21702
Female3773 ± 119
DiagnosisFemoral neck fracture3749 ± 133
Femoral head necrosis21738

Group III (n = 2): pronase E + collagenase IIAge (y)60–6911291
70–791170
SexMale11291
Female1170
DiagnosisFemoral neck fracture2730

Donor patient characteristics and wet weight of harvested cartilage for each treatment group

Group No. (cartilage treatment; sample size)Patient No.Age (y)SexDiagnosisWet weight of cartilage (mg) (mean ± SD)
I (Pronase E + Collagenase IA; n = 11)176FLeft femoral neck fracture4396 ± 1546
262MRight femoral head necrosis
362FRight femoral head necrosis
481FRight femoral neck fracture
577MLeft femoral neck fracture
662FLeft femoral neck fracture
761FRight knee osteoarthritis
874MLeft femoral neck fracture
975FLeft knee osteoarthritis
1070FRight hip osteoarthritis
1179FLeft femoral neck fracture

II (Collagenase II; n = 5)1283FLeft femoral neck fracture2854 ± 876.5
1367FRight femoral head necrosis
1455MRight femoral head necrosis
1572FRight femoral neck fracture
1687MLeft femoral neck fracture

III (Pronase E + Collagenase II; n = 2)1764MLeft femoral neck fracture2640
1872FLeft femoral neck fracture
DOI: https://doi.org/10.2478/abm-2021-0011 | Journal eISSN: 1875-855X | Journal ISSN: 1905-7415
Language: English
Page range: 91 - 99
Published on: Apr 30, 2021
Published by: Chulalongkorn University
In partnership with: Paradigm Publishing Services
Publication frequency: 6 issues per year

© 2021 Liuliu Xiong, Meng Cui, Ziye Zhou, Minchen Wu, Quanming Wang, Haiyan Song, Lei Ding, published by Chulalongkorn University
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.