CN1850303A - Composite skin containing self-body epidermic cells and variant epidermic cells - Google Patents
Composite skin containing self-body epidermic cells and variant epidermic cells Download PDFInfo
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- CN1850303A CN1850303A CN 200610027043 CN200610027043A CN1850303A CN 1850303 A CN1850303 A CN 1850303A CN 200610027043 CN200610027043 CN 200610027043 CN 200610027043 A CN200610027043 A CN 200610027043A CN 1850303 A CN1850303 A CN 1850303A
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- composite skin
- cell
- allosome
- body surface
- chrotoplast
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Abstract
The present invention relates to the field of medical wound repair technology, and provides a composite skin containing autogenous epidermic cells and heterogenous epidermic cells for making deep wound repair. Said invention is characterized by that it adopts conventional cell culture technique to make autogenous epidermic cells and heterogenous epidermic cell be mixed, then inoculated in the dermal succedaneum surface, then utilizes external culture to form the invented composite skin.
Description
Technical field
The present invention relates to medical science wound repair technical field, particularly a kind of Composite Skin of containing that is used for degree of depth skin injury wound repair from body surface chrotoplast and allosome epidermis cell.
Background technology
For solving extensive deep burn patient Pi Yuan problem of shortage, the Composite Skin that external structure contains epidermis cell is used for transplanting the focus that has become the wound repair field.The conventional at present method for preparing Composite Skin is: when the patient performs the operation for the first time, get fritter skin sample (2cm
2～4cm
2), separation is from the body surface chrotoplast, cultivate amplification, be inoculated in dermal substitute (as acellular dermal, spongy collagem membrane etc.) surface then, continuation is cultivated, and forms the Composite Skin that contains monolayer or multiple layer epidermis cell, with the deep burn wound of Composite skin in the underwent operative processing, can form new skin histology, thus wound repairing.But make up Composite Skin in this way, to reach certain amplification times from the body surface chrotoplast, could be used for clinical effectively, therefore often need 3～4 weeks of In vitro culture even longer time, can not in time be applied to clinical, especially the patient who lacks for wound surface Da Pi source electrode degree not only obtains and competently is restricted from the body surface chrotoplast, and very long cell culture period has seriously limited the clinical practice of Composite Skin.
Summary of the invention
The purpose of this invention is to provide a kind ofly except containing the Composite Skin that from the body surface chrotoplast, also contains the allosome epidermis cell, saving, and shorten the cells in vitro incubation time, as early as possible Composite Skin is applied to the patient, help wound healing from body skin source.
The present invention improves used clinically only containing from the Composite Skin of body surface chrotoplast at present, is about to inoculate on the dermal substitute surface after body surface chrotoplast and the mixing of allosome epidermis cell make up Composite Skin.The allosome epidermis cell can adopt routine techniques to cultivate, increase also frozen standby in liquid nitrogen.When making up Composite Skin, an amount of allosome epidermis cell can thaw temporarily, recover, with obtain after the body surface chrotoplast mixes, be inoculated in dermal substitute (as acellular dermal, spongy collagem membrane etc.) surface clinically commonly used, the cell density of inoculation is 2.5-5 * 10
5Individual/cm
2, in cell culture fluid, to cultivate 1-2 week routinely, cell culture fluid can be selected from epidermis cell culture fluid such as DMEM or SFM, can form the Composite Skin that contains monolayer or multiple layer epidermal sheet.During use, this Composite Skin is implanted wound surface such as deep burn that underwent operative handles well or chronic skin ulcer routinely.Since the allosome epidermis cell in the Composite Skin after transplanting can be repelled by body gradually and by periphery from body surface chrotoplast creeping substitution, therefore this Composite Skin had both made exposed wound surface be covered treatment early, can avoid again because of importing the potential hazard that the allosome epidermis cell may cause.
Composite Skin of the present invention compares with the simple Composite Skin of using always clinically at present from the body surface chrotoplast that contains, have significant advantage: Composite Skin of the present invention contains the allosome epidermis cell, the allosome epidermis cell can cultivate in advance, frozen, can thaw at any time when needing, recover and make up Composite Skin from body surface chrotoplast Mixed culture, the epidermis cell In vitro culture time can obviously be shortened like this, not only saved from body surface skin amount, and can in time be applied to clinical.Such as will be from body surface chrotoplast and allosome epidermis cell by 1: 1 mixed, with simple cultivate to transplant from the body surface chrotoplast compare, can save 50% from body surface skin amount, reduce by 50% incubation time.If by 1: 4 mixed, then can save 80% from body surface skin amount, reduce by about 4/5ths In vitro culture time, the large-area burns patient's who lacks for skin source electrode degree treatment is significant.
The specific embodiment
The construction method of Composite Skin of the present invention is:
1. the allosome epidermis cell is cultivated: get discarded skin histology of normal person such as foreskin, adopt conventional epidermis cell culture technique, the separating table cortex is prepared into single epidermal cell suspension, presses 2.5-5 * 10
5Individual/cm
2Cell density is inoculated in the culture bottle, places 37 ℃ of incubators to cultivate with perfect form DMEM culture fluid, goes down to posterity, increases when epidermis cell reaches 70%～80% fusion.When the allosome epidermis cell reaches some, frozen standby in liquid nitrogen.
2. cultivate from the body surface chrotoplast: cut patient's fritter skin sample such as 2cm
2～4cm
2, adopt conventional epidermis cell culture technique, the separating table cortex, prepare single epidermal cell suspension, be inoculated in culture bottle and cultivate, increase (specifically cultivating),, place culture fluid stand-by when when the body surface chrotoplast can satisfy the quantity of application need with the allosome epidermis cell.
3. structure Composite Skin: the allosome epidermis cell that thaws, recovers an amount of, with proper proportion with mix from the body surface chrotoplast after be inoculated in dermal substitute (as acellular dermal, spongy collagem membrane etc.) surface clinically commonly used routinely, add perfect form DMEM culture fluid through In vitro culture 1-2 week, change liquid once in per 2～3 days, and formed the Composite Skin that contains monolayer or multiple layer epidermal sheet.
The present invention contains the Composite Skin from body surface chrotoplast and allosome epidermis cell, and main uses has two kinds.First kind, be used for the reparation of skin injury wound surface such as extensive deep burn, hypertrophic cicatrix, chronic degree of depth skin ulcer as permanent Graftskin.Show through a large amount of animal (nude mice) experiments, to press 1: 1,1: 3,1: 5 mixed from body surface chrotoplast and allosome epidermis cell, be inoculated in acellular dermal substitute surface and cultivate back formation Composite Skin, face down with dermal substitute one then, implant holostrome skin injury wound surface, survival rate reaches 65% fully.After the Composite skin, the allosome epidermis cell prolongs with transplant time is ostracised gradually, and quilt is from body surface chrotoplast creeping substitution, and wound surface does not have obviously exposed.The visible basal layer of newborn skin, substrate upper strata and cuticular layer, epidermis and corium syndeton are normal, and arrangement of collagen fibers is regular in the corium.Second kind of purposes is used to promote the reparation of skin donor site, superficial burns wound surface as temporary covering.The Composite Skin that will contain from body surface chrotoplast and allosome epidermis cell faces down with cell, be affixed on wound surface, can serve as temporary covering, epidermis cell promotes remaining epithelium regeneration by iuntercellular contact, the release various kinds of cell factor, quickens the healing of skin donor site, superficial burns wound surface or chronic skin ulcer wound surface.
Embodiment 1 preparation allosome epidermis cell: get the discarded skin histology 4cm of normal person
2One, adopt conventional epidermis cell culture technique, the separating table cortex is prepared into single epidermal cell suspension, by 5 * 10
5Individual/cm
2Cell density is inoculated in the culture bottle, places 37 ℃ of incubators to cultivate for 1 week with perfect form DMEM culture fluid, and epidermis cell reaches 70%～80% and merges, and goes down to posterity, increases.When the allosome epidermis cell reaches 1 * 10
8Individual/during ml, frozen standby in liquid nitrogen after the packing.
Embodiment 2 preparation Composite Skin
(1) cultivates from the body surface chrotoplast: cut patient's fritter skin sample 2cm
2, adopting conventional epidermis cell culture technique, the separating table cortex prepares single epidermal cell suspension, is inoculated in culture bottle and cultivates, increases (specifically cultivating with the allosome epidermis cell), reaches 1 * 10 from the body surface chrotoplast
8Individual/as during ml, to place culture fluid standby.
(2) preparation Composite Skin: will frozen allosome epidermis cell thaw recover after, with from the body surface chrotoplast with 1: 1 mixed, be inoculated in the acellular dermal rack surface routinely, cell density is 5 * 10
5Individual/cm
2, in perfect form DMEM culture fluid, cultivated for 1 week down for 37 ℃, changed liquid 1 time in per 2 days, promptly form Composite Skin.
Embodiment 3
Preparation is during Composite Skin, and allosome epidermis cell and be 1: 5 from the ratio of body surface chrotoplast is by 4 * 10
5Individual/cm
2Cell density is inoculated in the acellular dermal rack surface, and culture fluid is DMEM, and all the other conditions are cultivated and promptly formed Composite Skin in 10 days with embodiment 2.
Embodiment 4
Preparation is during Composite Skin, and allosome epidermis cell and be 1: 1 from the ratio of body surface chrotoplast is by 5 * 10
5Individual/cm
2Cell density is inoculated in spongy collagem membrane surface, and all the other are cultivated and 1 week promptly formed Composite Skin with embodiment 2.
Embodiment 5
Preparation is during Composite Skin, and allosome epidermis cell and be 1: 3 from the ratio of body surface chrotoplast is by 2.5 * 10
5Individual/cm
2Cell density is inoculated in spongy collagem membrane surface, and all the other are cultivated and 2 weeks promptly formed Composite Skin with embodiment 2.
Claims (6)
1. Composite Skin that contains from body surface chrotoplast and allosome epidermis cell by dermal substitute be covered in its surperficial monolayer or a multiple layer epidermal sheet formed, is characterized in that containing in the epidermal sheet from body surface chrotoplast and allosome epidermis cell.
2. the preparation method of the described Composite Skin of claim 1 is characterized in that being inoculated in dermal substitute surface commonly used clinically with what prepare after body surface chrotoplast and the mixing of allosome epidermis cell, and the cell density of inoculation is 2.5 * 10
5～5 * 10
5Individual/cm
2, in cell culture fluid, cultivated for 1～2 week routinely, Composite Skin.
3. by the described Composite Skin preparation method of claim 2, it is characterized in that the mixed proportion from body surface chrotoplast and allosome epidermis cell is 1: 1～1: 5.
4. by claim 2 or 3 described Composite Skin preparation methoies, it is characterized in that said dermal substitute is acellular dermal substitute or spongy collagem membrane.
5. by claim 2 or 3 described Composite Skin preparation methoies, it is characterized in that said cell culture fluid is the DMEM culture fluid.
6. by the described Composite Skin preparation method of claim 4, it is characterized in that said cell culture fluid is the DMEM culture fluid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610027043 CN1850303A (en) | 2006-05-30 | 2006-05-30 | Composite skin containing self-body epidermic cells and variant epidermic cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 200610027043 CN1850303A (en) | 2006-05-30 | 2006-05-30 | Composite skin containing self-body epidermic cells and variant epidermic cells |
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| Publication Number | Publication Date |
|---|---|
| CN1850303A true CN1850303A (en) | 2006-10-25 |
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ID=37131814
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 200610027043 Pending CN1850303A (en) | 2006-05-30 | 2006-05-30 | Composite skin containing self-body epidermic cells and variant epidermic cells |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106957817A (en) * | 2017-02-21 | 2017-07-18 | 安徽安龙基因医学检验所有限公司 | A kind of construction method for being used to repair the cytoskeleton without the meniscus injury of Xue Yun areas |
-
2006
- 2006-05-30 CN CN 200610027043 patent/CN1850303A/en active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106957817A (en) * | 2017-02-21 | 2017-07-18 | 安徽安龙基因医学检验所有限公司 | A kind of construction method for being used to repair the cytoskeleton without the meniscus injury of Xue Yun areas |
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Open date: 20061025 |